Detergent composition comprising GH9 endoglucanase variants I

ABSTRACT

The present disclosure relates to a detergent composition comprising endoglucanase variants and methods for use of said compositions.

CROSS-REFERENCE TO RELATED APPLICATION

This application is a U.S. National-Stage entry under 35 U.S.C. § 371based on International Application No. PCT/EP2017/071297, filed Aug. 24,2017, which was published under PCT Article 21(2) and which claimspriority to German Application No. 10 2016 215 867.8, filed Aug. 24,2016, which are all hereby incorporated in their entirety by reference.

REFERENCE TO A JOINT RESEARCH AGREEMENT

The embodiments claimed in the present application were made under ajoint research agreement between Henkel AG & Co. KGaA and Novozymes A/S.

REFERENCE TO A SEQUENCE LISTING

This application contains a Sequence Listing in computer readable form,which is incorporated herein by reference.

TECHNICAL FIELD

The present disclosure relates to a detergent composition, such aslaundry compositions and dish wash compositions, including hand wash andautomatic dish wash compositions, comprising novel GH9 endoglucanasevariants exhibiting alterations relative to the parent GH9 endoglucanasein one or more properties including: detergent stability (e.g. improvedstability in a detergent composition, e.g. in the presence of achelator, e.g. EDTA or citrate) and/or storage stability (e.g. improvedstorage stability in a detergent composition, e.g. in the presence of achelator, e.g. EDTA or citrate). The present disclosure further relatesto detergent compositions comprising novel GH9 endoglucanase variantshaving activity on xanthan gum pretreated with xanthan lyase. Thedisclosure also relates to methods for producing and using thecompositions of the disclosure. Variants described herein areparticularly suitable for use in cleaning processes and detergentcompositions.

BACKGROUND

Xanthan gum is a polysaccharide derived from the bacterial coat ofXanthomonas campestris. It is produced by the fermentation of glucose,sucrose, or lactose by the Xanthomonas campestris bacterium. After afermentation period, the polysaccharide is precipitated from a growthmedium with isopropyl alcohol, dried, and ground into a fine powder.Later, it is added to a liquid medium to form the gum. Xanthan gum is anatural polysaccharide including different sugars which are connected byseveral different bonds, such as β-D-mannosyl-β-D-1,4-glucuronosyl bondsand β-D-glucosyl-β-D-1,4-glucosyl bonds. Xanthan gum is at least partlysoluble in water and forms highly viscous solutions or gels. Completeenzymatic degradation of xanthan gum requires several enzymaticactivities including xanthan lyase activity and endo-β-1,4-glucanaseactivity. Xanthan lyases are enzymes that cleave theβ-D-mannosyl-β-D-1,4-glucuronosyl bond of xanthan and have beendescribed in the literature. Xanthan lyases are known in the art, e.g.two xanthan lyases have been isolated from Paenibacillus alginolyticusXL-1 (e.g. Ruijssenaars et al. (1999) ‘A pyruvated mannose-specificxanthan lyase involved in xanthan degradation by Paenibacillusalginolyticus XL-1’, Appl. Environ. Microbiol. 65(6): 2446-2452, andRuijssenaars et al. (2000), ‘A novel gene encoding xanthan lyase ofPaenibacillus alginolyticus strain XL-1’, Appl. Environ. Microbiol.66(9): 3945-3950). Glycoside hydrolases are enzymes that catalyse thehydrolysis of the glycosyl bond to release smaller sugars. There areover 100 classes of glycoside hydrolases which have been classified, seeHenrissat et al. (1991) ‘A classification of glycosyl hydrolases basedon amino-acid sequence similarities’, J. Biochem. 280: 309-316 and theUniprot website at www.cazy.org. The glycoside hydrolase family 9 (GH9)includes over 70 different enzymes that are mostly endo-glucanases (EC3.2.1.4), cellobiohydrolases (EC 3.2.1.91), β-glucosidases (EC 3.2.1.21)and exo-β-glucosaminidase (EC 3.2.1.165). In recent years xanthan gumhas been used as an ingredient in many consumer products including foods(e.g. as thickening agent in salad dressings and dairy products) andcosmetics (e.g. as stabilizer and thickener in toothpaste and make-up,creams and lotions to prevent ingredients from separating and to providethe right texture of the product). Further xanthan gum has found use inthe oil industry as an additive to regulate the viscosity of drillingfluids etc. The widespread use of xanthan gum has led to a desire todegrade solutions, gels or mixtures containing xanthan gum therebyallowing easier removal of the byproducts. Endoglucanases and xanthanlyases for the degradation of xanthan gum and the use of such enzymesfor cleaning purposes, such as the removal of xanthan gum containingstains, and in the drilling and oil industries are known in the art,e.g. WO2013167581A1.

The known xanthan endoglucanase having SEQ ID NO: 2 was found to besensitive to the presence of detergents with chelators. To improve theapplicability and/or cost and/or the performance of such enzymes thereis an ongoing search for variants with altered properties, such asincreased stability, e.g. improved stability in a detergent composition,e.g. in the presence of a chelator, e.g. EDTA or citrate, etc. However,mutagenesis of large enzymes followed by purification and functionalanalysis of mutant libraries can be very expensive and laborious.

BRIEF SUMMARY

The instant disclosure provides a detergent composition including anendoglucanase variant, comprising an alteration at one or more positionsin one or more chelator-induced instability region selected from thegroup of:

-   -   i) region 7 corresponding to amino acids 612 to 660 of SEQ ID        NO: 2    -   ii) region 1 corresponding to amino acids 95 to 105 of SEQ ID        NO: 2,    -   iii) region 2 corresponding to amino acids 115 to 138 of SEQ ID        NO: 2,    -   iv) region 3 corresponding to amino acids 210 to 251 of SEQ ID        NO: 2,    -   v) region 4 corresponding to amino acids 267 to 301 of SEQ ID        NO: 2,    -   vi) region 5 corresponding to amino acids 339 to 361 of SEQ ID        NO: 2,    -   vii) region 6 corresponding to amino acids 547 to 595 of SEQ ID        NO: 2,    -   viii) region 8 corresponding to amino acids 806 to 828 of SEQ ID        NO: 2, and    -   ix) region 9 corresponding to amino acids 839 to 1042 of SEQ ID        NO: 2,    -   wherein said variant has at least about 60% and less than about        100% sequence identity to SEQ ID NO: 2.

DETAILED DESCRIPTION

The following detailed description is merely exemplary in nature and isnot intended to limit the disclosure or the application and uses of thesubject matter as described herein. Furthermore, there is no intentionto be bound by any theory presented in the preceding background or thefollowing detailed description.

Since the known xanthan endoglucanase having SEQ ID NO: 2 is a largeenzyme (>1000 residues), it is difficult and expensive to randomlytarget its properties for improvement of, e.g., stability in a detergentcomposition, e.g., in the presence of a chelator.

In some aspects, the present disclosure identifies chelator-inducedinstability regions in the protein sequence/structure of the knownxanthan endoglucanase having SEQ ID NO: 2 that are affected when themolecule is incubated in a buffer with EDTA, and therefore provides animportant guidance on where to mutate an endoglucanase in order tostabilize the molecule in a detergent, e.g., detergent compositioncomprising a chelator.

In some aspects, the present disclosure relates to a detergentcomposition comprising an endoglucanase variant, comprising analteration (e.g., a substitution, deletion or insertion) at one or morepositions in a chelator-induced instability region selected from thegroup of: region 1 corresponding to amino acids 95 to 105 of SEQ ID NO:2, region 2 corresponding to amino acids 115 to 138 of SEQ ID NO: 2,region 3 corresponding to amino acids 210 to 251 of SEQ ID NO: 2, region4 corresponding to amino acids 267 to 301 of SEQ ID NO: 2, region 5corresponding to amino acids 339 to 361 of SEQ ID NO: 2, region 6corresponding to amino acids 547 to 595 of SEQ ID NO: 2, region 7corresponding to amino acids 612 to 660 of SEQ ID NO: 2, region 8corresponding to amino acids 806 to 828 of SEQ ID NO: 2, and region 9corresponding to amino acids 839 to 1042 of SEQ ID NO: 2.

In some aspects, the present disclosure relates to a detergentcomposition comprising an endoglucanase variant, comprising analteration (e.g., a substitution, deletion or insertion) at one or morepositions in a region selected from the group of: region 1 correspondingto amino acids 95 to 105 of SEQ ID NO: 2, region 2 corresponding toamino acids 115 to 138 of SEQ ID NO: 2, region 3 corresponding to aminoacids 210 to 251 of SEQ ID NO: 2, region 4 corresponding to amino acids267 to 301 of SEQ ID NO: 2, region 5 corresponding to amino acids 339 to361 of SEQ ID NO: 2, region 6 corresponding to amino acids 547 to 595 ofSEQ ID NO: 2, region 7 corresponding to amino acids 612 to 660 of SEQ IDNO: 2, region 8 corresponding to amino acids 806 to 828 of SEQ ID NO: 2,and region 9 corresponding to amino acids 839 to 1042 of SEQ ID NO: 2,wherein said variant has at least about 60% and less than about 100%sequence identity to SEQ ID NO: 2; preferably said endoglucanase varianthas activity on xanthan gum pre-treated with xanthan lyase.

In some aspects, the chelator-induced instability region of a parentendoglucanase (e.g., SEQ ID NO: 2) has one or more of the followingfeatures: in the presence of a chelator it is relatively lessconformationally stable than one or more or all of its adjacent regions;and/or in the presence of a chelator it is relatively more exposed tosaid chelator than one or more or all of its adjacent regions; and/or inthe presence of a chelator it is relatively more accessible to saidchelator than one or more or all of its adjacent regions; and/or in thepresence of a chelator it is relatively more conformationally dynamicthan one or more or all of its adjacent regions; and/or in the presenceof a chelator it is relatively more receptive to deuterium incorporationthan one or more or all of its adjacent regions. In the presentdisclosure, when referring to “relatively” it means that the features ofa given region as indicated above are based on observed differencesbetween its features and/or properties in the presence of a chelator andits features and/or properties in the absence of a chelator (i.e. inorder to determine the impact of a chelator, each region is compared toitself in the absence of said chelator and any changes are determinedrelative to its native features/properties in the absence of a chelator)(see also Example 1).

In some aspects, the present disclosure relates to a detergentcomposition comprising an endoglucanase variant having at least about61%, at least about 62%, at least about 63%, at least about 64%, atleast about 65%, at least about 66%, at least about 67%, at least about68%, at least about 69%, at least about 70%, at least about 71%, atleast about 72%, at least about 73%, at least about 74%, at least about75%, at least about 76%, at least about 77%, at least about 78%, atleast about 79%, at least about 80%, at least about 81%, at least about82%, at least about 83%, at least about 84%, at least about 85%, atleast about 86%, at least about 87%, at least about 88%, at least about89%, at least about 90%, at least about 91%, at least about 92%, atleast about 93%, at least about 94%, at least about 95%, at least about96%, at least about 97%, at least about 98%, or at least about 99%sequence identity to SEQ ID NO: 2.

In some aspects, the present disclosure relates to a detergentcomposition comprising an endoglucanase variant, comprising analteration (e.g., a substitution, deletion or insertion) at one or morepositions in a region selected from the group of:

i) region 1 corresponding to amino acids 95 to 105 of SEQ ID NO: 2,e.g., said alteration at one or more positions selected from the groupof positions: 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, whereinsaid positions correspond to amino acid positions of SEQ ID NO: 2 (e.g.,using the numbering of SEQ ID NO: 2),

ii) region 2 corresponding to amino acids 115 to 138 of SEQ ID NO: 2,e.g., said alteration at one or more positions selected from the groupof positions: 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125,126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, whereinsaid positions correspond to amino acid positions of SEQ ID NO: 2 (e.g.,using the numbering of SEQ ID NO: 2),

iii) region 3 corresponding to amino acids 210 to 251 of SEQ ID NO: 2,e.g., said alteration at one or more positions selected from the groupof positions: 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220,221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234,235, 236, 237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248,249, 250, 251, wherein said positions correspond to amino acid positionsof SEQ ID NO: 2 (e.g., using the numbering of SEQ ID NO: 2),

iv) region 4 corresponding to amino acids 267 to 301 of SEQ ID NO: 2,e.g., said alteration at one or more positions selected from the groupof positions: 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277,278, 279, 280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291,292, 293, 294, 295, 296, 297, 298, 299, 300, 301, wherein said positionscorrespond to amino acid positions of SEQ ID NO: 2 (e.g., using thenumbering of SEQ ID NO: 2),

v) region 5 corresponding to amino acids 339 to 361 of SEQ ID NO: 2,e.g., said alteration at one or more positions selected from the groupof positions: 339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349,350, 351, 352, 353, 354, 355, 356, 357, 358, 359, 360, 361, wherein saidpositions correspond to amino acid positions of SEQ ID NO: 2 (e.g.,using the numbering of SEQ ID NO: 2),

vi) region 6 corresponding to amino acids 547 to 595 of SEQ ID NO: 2,e.g., said alteration at one or more positions selected from the groupof positions: 547, 548, 549, 550, 551, 552, 553, 554, 555, 556, 557,558, 559, 560, 561, 562, 563, 564, 565, 566, 567, 568, 569, 570, 571,572, 573, 574, 575, 576, 577, 578, 579, 580, 581, 582, 583, 584, 585,586, 587, 588, 589, 590, 591, 592, 593, 594, 595, wherein said positionscorrespond to amino acid positions of SEQ ID NO: 2 (e.g., using thenumbering of SEQ ID NO: 2),

vii) region 7 corresponding to amino acids 612 to 660 of SEQ ID NO: 2,e.g., said alteration at one or more positions selected from the groupof positions: 612, 613, 614, 615, 616, 617, 618, 619, 620, 621, 622,623, 624, 625, 626, 627, 628, 629, 630, 631, 632, 633, 634, 635, 636,637, 638, 639, 640, 641, 642, 643, 644, 645, 646, 647, 648, 649, 650,651, 652, 653, 654, 655, 656, 657, 658, 659, 660, wherein said positionscorrespond to amino acid positions of SEQ ID NO: 2 (e.g., using thenumbering of SEQ ID NO: 2),

viii) region 8 corresponding to amino acids 806 to 828 of SEQ ID NO: 2,e.g., said alteration at one or more positions selected from the groupof positions: 806, 807, 808, 809, 810, 811, 812, 813, 814, 815, 816,817, 818, 819, 820, 821, 822, 823, 824, 825, 826, 827, 828, wherein saidpositions correspond to amino acid positions of SEQ ID NO: 2 (e.g.,using the numbering of SEQ ID NO: 2), and

ix) region 9 corresponding to amino acids 839 to 1042 of SEQ ID NO: 2,e.g., said alteration at one or more positions selected from the groupof positions: 839, 840, 841, 842, 843, 844, 845, 846, 847, 848, 849,850, 851, 852, 853, 854, 855, 856, 857, 858, 859, 860, 861, 862, 863,864, 865, 866, 867, 868, 869, 870, 871, 872, 873, 874, 875, 876, 877,878, 879, 880, 881, 882, 883, 884, 885, 886, 887, 888, 889, 890, 891,892, 893, 894, 895, 896, 897, 898, 899, 900, 901, 902, 903, 904, 905,906, 907, 908, 909, 910, 911, 912, 913, 914, 915, 916, 917, 918, 919,920, 921, 922, 923, 924, 925, 926, 927, 928, 929, 930, 931, 932, 933,934, 935, 936, 937, 938, 939, 940, 941, 942, 943, 944, 945, 946, 947,948, 949, 950, 951, 952, 953, 954, 955, 956, 957, 958, 959, 960, 961,962, 963, 964, 965, 966, 967, 968, 969, 970, 971, 972, 973, 974, 975,976, 977, 978, 979, 980, 981, 982, 983, 984, 985, 986, 987, 988, 989,990, 991, 992, 993, 994, 995, 996, 997, 998, 999, 1000, 1001, 1002,1003, 1004, 1005, 1006, 1007, 1008, 1009, 1010, 1011, 1012, 1013, 1014,1015, 1016, 1017, 1018, 1019, 1020, 1021, 1022, 1023, 1024, 1025, 1026,1027, 1028, 1029, 1030, 1031, 1032, 1033, 1034, 1035, 1036, 1037, 1038,1039, 1040, 1041, 1042, wherein said positions correspond to amino acidpositions of SEQ ID NO: 2 (e.g., using the numbering of SEQ ID NO: 2).

In some aspects, the present disclosure relates to a detergentcomposition comprising an endoglucanase variant, comprising analteration (e.g., a substitution, deletion or insertion) at one or morepositions in:

a) one or more regions selected from the group of:

-   -   i) region 1 corresponding to amino acids 95 to 105 of SEQ ID NO:        2, e.g., said alteration at one or more positions selected from        the group of positions: 95, 96, 97, 98, 99, 100, 101, 102, 103,        104, 105, wherein said positions correspond to amino acid        positions of SEQ ID NO: 2 (e.g., using the numbering of SEQ ID        NO: 2),    -   ii) region 2 corresponding to amino acids 115 to 138 of SEQ ID        NO: 2, e.g., said alteration at one or more positions selected        from the group of positions: 115, 116, 117, 118, 119, 120, 121,        122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134,        135, 136, 137, 138, wherein said positions correspond to amino        acid positions of SEQ ID NO: 2 (e.g., using the numbering of SEQ        ID NO: 2),    -   iii) region 3 corresponding to amino acids 210 to 251 of SEQ ID        NO: 2, e.g., said alteration at one or more positions selected        from the group of positions: 210, 211, 212, 213, 214, 215, 216,        217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229,        230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 242,        243, 244, 245, 246, 247, 248, 249, 250, 251, wherein said        positions correspond to amino acid positions of SEQ ID NO: 2        (e.g., using the numbering of SEQ ID NO: 2),    -   iv) region 4 corresponding to amino acids 267 to 301 of SEQ ID        NO: 2, e.g., said alteration at one or more positions selected        from the group of positions: 267, 268, 269, 270, 271, 272, 273,        274, 275, 276, 277, 278, 279, 280, 281, 282, 283, 284, 285, 286,        287, 288, 289, 290, 291, 292, 293, 294, 295, 296, 297, 298, 299,        300, 301, wherein said positions correspond to amino acid        positions of SEQ ID NO: 2 (e.g., using the numbering of SEQ ID        NO: 2),    -   v) region 5 corresponding to amino acids 339 to 361 of SEQ ID        NO: 2, e.g., said alteration at one or more positions selected        from the group of positions: 339, 340, 341, 342, 343, 344, 345,        346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357, 358,        359, 360, 361, wherein said positions correspond to amino acid        positions of SEQ ID NO: 2 (e.g., using the numbering of SEQ ID        NO: 2),    -   vi) region 6 corresponding to amino acids 547 to 595 of SEQ ID        NO: 2, e.g., said alteration at one or more positions selected        from the group of positions: 547, 548, 549, 550, 551, 552, 553,        554, 555, 556, 557, 558, 559, 560, 561, 562, 563, 564, 565, 566,        567, 568, 569, 570, 571, 572, 573, 574, 575, 576, 577, 578, 579,        580, 581, 582, 583, 584, 585, 586, 587, 588, 589, 590, 591, 592,        593, 594, 595, wherein said positions correspond to amino acid        positions of SEQ ID NO: 2 (e.g., using the numbering of SEQ ID        NO: 2),    -   vii) region 7 corresponding to amino acids 612 to 660 of SEQ ID        NO: 2, e.g., said alteration at one or more positions selected        from the group of positions: 612, 613, 614, 615, 616, 617, 618,        619, 620, 621, 622, 623, 624, 625, 626, 627, 628, 629, 630, 631,        632, 633, 634, 635, 636, 637, 638, 639, 640, 641, 642, 643, 644,        645, 646, 647, 648, 649, 650, 651, 652, 653, 654, 655, 656, 657,        658, 659, 660, wherein said positions correspond to amino acid        positions of SEQ ID NO: 2 (e.g., using the numbering of SEQ ID        NO: 2),    -   viii) region 8 corresponding to amino acids 806 to 828 of SEQ ID        NO: 2, e.g., said alteration at one or more positions selected        from the group of positions: 806, 807, 808, 809, 810, 811, 812,        813, 814, 815, 816, 817, 818, 819, 820, 821, 822, 823, 824, 825,        826, 827, 828, wherein said positions correspond to amino acid        positions of SEQ ID NO: 2 (e.g., using the numbering of SEQ ID        NO: 2), and    -   ix) region 9 corresponding to amino acids 839 to 1042 of SEQ ID        NO: 2, e.g., said alteration at one or more positions selected        from the group of positions: 839, 840, 841, 842, 843, 844, 845,        846, 847, 848, 849, 850, 851, 852, 853, 854, 855, 856, 857, 858,        859, 860, 861, 862, 863, 864, 865, 866, 867, 868, 869, 870, 871,        872, 873, 874, 875, 876, 877, 878, 879, 880, 881, 882, 883, 884,        885, 886, 887, 888, 889, 890, 891, 892, 893, 894, 895, 896, 897,        898, 899, 900, 901, 902, 903, 904, 905, 906, 907, 908, 909, 910,        911, 912, 913, 914, 915, 916, 917, 918, 919, 920, 921, 922, 923,        924, 925, 926, 927, 928, 929, 930, 931, 932, 933, 934, 935, 936,        937, 938, 939, 940, 941, 942, 943, 944, 945, 946, 947, 948, 949,        950, 951, 952, 953, 954, 955, 956, 957, 958, 959, 960, 961, 962,        963, 964, 965, 966, 967, 968, 969, 970, 971, 972, 973, 974, 975,        976, 977, 978, 979, 980, 981, 982, 983, 984, 985, 986, 987, 988,        989, 990, 991, 992, 993, 994, 995, 996, 997, 998, 999, 1000,        1001, 1002, 1003, 1004, 1005, 1006, 1007, 1008, 1009, 1010,        1011, 1012, 1013, 1014, 1015, 1016, 1017, 1018, 1019, 1020,        1021, 1022, 1023, 1024, 1025, 1026, 1027, 1028, 1029, 1030,        1031, 1032, 1033, 1034, 1035, 1036, 1037, 1038, 1039, 1040,        1041, 1042, wherein said positions correspond to amino acid        positions of SEQ ID NO: 2 (e.g., using the numbering of SEQ ID        NO: 2); and/or

b) one or more adjacent regions, said adjacent region being selectedfrom the group of:

-   -   i′) region 10 corresponding to amino acids 1 to 94 of SEQ ID NO:        2,    -   ii′) region 11 corresponding to amino acids 106 to 114 of SEQ ID        NO: 2,    -   iii′) region 12 corresponding to amino acids 139 to 209 of SEQ        ID NO: 2,    -   iv′) region 13 corresponding to amino acids 252 to 266 of SEQ ID        NO: 2,    -   v′) region 14 corresponding to amino acids 302 to 338 of SEQ ID        NO: 2,    -   vi′) region 15 corresponding to amino acids 362 to 546 of SEQ ID        NO: 2,    -   vii′) region 16 corresponding to amino acids 596 to 611 of SEQ        ID NO: 2,    -   viii′) region 17 corresponding to amino acids 661 to 805 of SEQ        ID NO: 2,    -   ix′) region 18 corresponding to amino acids 829 to 838 of SEQ ID        NO: 2, and    -   x′) region 19 corresponding to amino acids 1043 to 1055 of SEQ        ID NO: 2.

In some aspects, the present disclosure relates to a detergentcomposition comprising an endoglucanase variant having an alteration(e.g., a substitution, deletion or insertion) at one or more positionsselected from the group of positions: 4, 17, 18, 20, 51, 53, 55, 56, 60,63, 71, 79, 87, 92, 99, 120, 125, 126, 130, 137, 182, 186, 189, 192,213, 216, 221, 226, 228, 230, 231, 232, 233, 235, 240, 243, 247, 249,278, 279, 281, 283, 285, 289, 292, 294, 298, 302, 311, 313, 333, 346,353, 358, 386, 387, 388, 390, 403, 408, 410, 416, 441, 448, 451, 471,472, 476, 489, 507, 512, 515, 538, 555, 556, 557, 558, 559, 560, 561,562, 563, 564, 567, 568, 570, 575, 578, 579, 580, 581, 583, 589, 590,591, 592, 593, 595, 598, 599, 602, 603, 605, 607, 609, 616, 627, 630,631, 635, 636, 638, 639, 640, 641, 642, 643, 644, 651, 676, 683, 688,690, 694, 698, 699, 706, 711, 713, 1719, 720, 744, 749, 754, 756, 760,781, 786, 797, 810, 811, 812, 815, 823, 824, 825, 827, 828, 833, 834,835, 837, 843, 848, 868, 869, 870, 871, 872, 873, 874, 880, 881, 883,884, 885, 887, 888, 890, 892, 894, 898, 905, 906, 912, 920, 921, 924,926, 927, 928, 932, 933, 934, 935, 937, 938, 939, 940, 941, 942, 943,946, 948, 950, 952, 953, 954, 956, 957, 960, 966, 971, 972, 980, 989,991, 994, 995, 998, 999, 1006, 1009, 1010, 1011, 1029, 1030, 1031, 1032,1035, 1037, 1038, 1040, 1041, 1042, 1044, 1045, and 1048, whereinnumbering is according to SEQ ID NO: 2.

In one alternative embodiment, the endoglucanase variant comprises analteration in one or more positions selected from the group of: 285,333, 353, 558, 633, 635, 638, 639, 994, 281, 563, 575, 921,558+559+560+561+562, 558, 559, 560, 561, 562 125, 126, 130, 213, 221,228, 230, 231, 232, 235, 240, 243, 249, 278, 292, 297, 346, 556, 564,565, 567, 568, 569, 570, 576, 578, 579, 580, 583, 589, 590, 591, 592,593, 616, 627, 630, 636, 641, 642, 643, 644, 651, 810, 811, 812, 815,823, 824, 825, 827, 843, 870, 871, 872, 873, 874, 881, 883, 884, 885,887, 894, 920, 932, 933, 934, 935, 937, 938, 939, 940, 941, 942, 943,950, 952, 953, 954, 960, 964, 966, 971, 974, 989, 991, 995, 998, 1006,1010, 1011, 1029, 1030, 1031, 1032, 1035, 1037, 1038, 1040, 1041, 1044,1045, 559+579, 564+579, 562+579, 559+579+99, 99, 559+579+281,281+559+579, 559+579+616, 559+579+636, 559+579+651, 559+579+948, 948,559+579+1009, 1009, 559+579+627, 579+921, 559+579+921, 99+579, 579+651,579+948, 579+1009, 559+579+934, 934, 559+579+921+934, 559+579+627,559+579+627+616, 559+579+627, 559+579+921+651, 559+579+921+627,559+579+921+636, 559+579+921+616, 559+579+921+636, 559+579+921+627+636,559+579+636+651, 559+579+616+651, 559+579+616+636, 559+579+616+921+934,559+579+651+627, 559+579+651+636, 559+579+651+627+636, 559+579+651+616,559+579+651+921+934, 636+934, 636+921, 636+627, 636+579, 638+934,638+921, 638+627, 638+579, 627+51, 51, 627+451, 451, 627+559, 627+579,579+934, 651+638, 570+651, 570+921, 570+627, 570+559, 570+579, 570+638,570+579, 570+638, 570+651, 570+636, 570+934, 570+638, 570+921, 570+627,570+559, 570+885, 885+934, 885+627, 559+579+636, 559+579+638,559+579+870, 559+579+560, 559+579+564, 559+579+570, 559+579+570,559+579+570, 559+579+570, 559+579+570, 559+579+570, 559+579+570,559+579+570, 559+570+579, 559+570+579, 559+570+579, 559+570+579,559+570+579, 559+570+579, 559+570+579, 559+570+579, 559+560+579,559+579+651, 559+579+651+934, 559+579+638, 559+579+921, 559+579+616+921,559+579+636, 559+579, 559+579, 559+579+921, 559+579+616, 638+934,627+636, 627+934, 570+579, 416+559+579+636, 128+559+579+627,128+559+579+636, and 579+636 of SEQ ID NO: 2.

In some aspects, the present disclosure relates to a detergentcomposition comprising an endoglucanase variant having an alteration(e.g., a substitution, deletion or insertion) at one or more positionsselected from the group of: N285G, W333L, T353D, N558NP, N558F, T633V,D635L, D635M, D635T, F638Y, T639D, G994N, and K281T, G563E, I575M,I575A, K921D, N558K+A559K+S560F+T561P+G562W, N558K, A559K, S560F, T561P,G562W and I125V, A126R, K130R, K213R, A221R, K228E, K228I, G230F, G230L,G230A, G230H, G230N, G230W, G230T, F231Y, F231N, V232R, V232G, H235D,N240Q, G243K, G243R, A249N, A278S, K281F, K281V, K281Y, K281H, K281Q,K281N, K281W, N285L, N285M, N285S, N285P, N285T, N285Y, N285H, N285K,N285D, N285W, N285R, T292F, T292L, T292I, T292V, T292S, T292P, T292Y,T292Q, T292N, T292K, T292D, T292G, F297L, A346H, G556S, N558D, N558M,N558Q, N558I, N558Y, N558H, A559N, A559F, A559M, A559P, A559Y, A559H,A559Q, A559D, A559R, A559G, A559I, A559S, S560P, S560K, S560G, S560D,T561P, T561E, T561Q, T561S, T561D, A564I, A564Y, A564H, A564Q, A564K,A564E, E565M, V567F, K568R, L569F, L569Y, L569D, L569E, P570F, P570L,P570I, P570M, P570V, P570S, P570T, P570A, P570Y, P570H, P570Q, P570N,P570K, P570E, P570W, P570R, P570G, I575D, I575E, I576F, I576M, I576P,D578R, Y579F, Y579W, V580L, D583M, Q589G, P590S, P590T, P590E, E591L,G592D, S593P, S593H, S593Q, S593N, S593K, S593D, S593E, S593R, S616D,K627L, K627M, K627V, K627S, K627T, K627Q, K627R, I630F, I630V, I630Y,D635A, D635P, D635N, D635K, D635E, D635G, D635W, S636L, S636M, S636A,S636H, S636Q, S636N, S636K, S636R, F638I, F638V, F638T, F638L, F638H,T639V, T639S, T639L, T639I, T639M, T639A, T639E, T639W, T639G, Y641E,S642T, S642N, N643D, N643H, N643T, T644F, A651P, S810R, A811S, V812F,V812I, V812M, V812W, V812R, N815V, N815Y, N815E, N815W, N815R, S823Q,A824T, T825N, T825W, T825A, T825D, V827I, V827M, V827S, T843V, D870F,D870L, D870I, D870M, D870V, D870S, D870T, D870Y, D870H, D870Q, D870N,D870K, D870E, D870W, D870R, D870G, P871F, P871L, P871I, P871M, P871V,P871S, P871T, P871A, P871Y, P871H, P871Q, T872S, T872F, T872A, T872Y,T872H, T872Q, T872N, T872K, T872D, T872E, T872W, T872R, T872G, D873K,D873E, T874V, T874S, T874P, T874A, T874H, T874Q, T874N, T874K, V881Q,T883K, Y884H, A885F, A885Q, A885N, T887L, T887I, T887S, T887H, T887R,K894E, N920D, K921R, K921E, T932A, N933V, N933S, Y934G, Y934M, Y934S,Y934A, Y934Q, Y934N, Y934E, Y934W, Y934R, T935W, A937F, A937V, A937S,A937T, A937Q, A937D, A937E, V938I, K939I, K939V, D940E, N941S, N941H,N941D, A942P, A942E, D943Y, D943H, R950V, R950H, R950N, F952S, F952W,N953Y, G954L, Y960F, A964N, A964C, N966P, N966C, G971A, Q974K, Q974C,Q989I, Q991L, Q991I, Q991M, Q991V, Q991T, Q991K, Q991C, S995I, S995V,S995Q, S995R, S995C, G998V, G998A, S1006T, S1006A, S1006K, S1006R,Y1010W, L1011M, L1011S, L1011A, L1011Q, L1011N, L1011D, L1011E, R1029N,F1030M, K1031I, K1031S, K1031T, K1031H, V1032G, K1035A, A1037E, A1037W,S1038L, S1038I, L1040N, L1040E, G1041F, L1044F, L1044S, L1044N, L1044W,P1045Q, P1045W, and A559N+Y579F, A559N, Y579F, A564E+Y579F, A564E,Y579F, A559N+Y579W, A559N, Y579W, G562P+Y579W, G562P, Y579W,A564D+Y579W, A564D, Y579W, A559N+Y579W+K99R, A559N, Y579W, K99R,A559N+Y579W+K281R, A559N, Y579W, K281R, K281R+A559N+Y579W, K281R, A559N,Y579W, A559N+Y579W+S616D, A559N, Y579W, S616D, A559N+Y579W+S636N, A559N,Y579W, S636N, A559N+Y579W+A651P, A559N, Y579W, A651P, A559N+Y579W+K948E,A559N, Y579W, K948E, A559N+Y579W+K1009E, A559N, Y579W, K1009E,A559N+Y579W+K627R, A559N, Y579W, K627R, Y579W+K921R, Y579W, K921R,A559N+Y579W+K921R, A559N, Y579W, K921R, K99R+Y579W, K99R, Y579W,Y579W+A651P, Y579W, A651P, Y579W+K948E, Y579W, K948E, Y579W+K1009E,Y579W, K1009E, A559N+Y579W+Y934G, A559N, Y579W, Y934G,A559N+Y579W+K921R+Y934G, A559N, Y579W, K921R, Y934G, A559N+Y579W+K627M,A559N, Y579W, K627M, A559N+Y579W+K627R+S616D, A559N, Y579W, K627R,S616D, A559N+Y579F+K627R, A559N, Y579F, K627R, A559N+Y579W+K921R+A651P,A559N, Y579W, K921R, A651P, A559N+Y579W+K921R+K627R, A559N, Y579W,K921R, K627R, A559N+Y579W+K921R+S636K, A559N, Y579W, K921R, S636K,A559N+Y579W+K921R+S616D, A559N, Y579W, K921R, S616D,A559N+Y579W+K921R+S636N, A559N, Y579W, K921R, S636N,A559N+Y579W+K921R+K627R+S636N, A559N, Y579W, K921R, K627R, S636N,A559N+Y579W+S636N+A651P, A559N, Y579W, S636N, A651P,A559N+Y579W+S616D+A651P, A559N, Y579W, S616D, A651P,A559N+Y579W+S616D+S636K, A559N, Y579W, S616D, S636K,A559N+Y579W+S616D+K921R+Y934G, A559N, Y579W, S616D, K921R, Y934G,A559N+Y579W+A651P+K627M, A559N, Y579W, A651P, K627M,A559N+Y579W+A651P+S636K, A559N, Y579W, A651P, S636K,A559N+Y579W+A651P+K627R+S636N, A559N, Y579W, A651P, K627R, S636N,A559N+Y579W+A651P+S616D, A559N, Y579W, A651P, S616D,A559N+Y579W+A651P+K921R+Y934G, A559N, Y579W, A651P, K921R, Y934G,S636N+Y934G, S636N, Y934G, S636N+K921R, S636N, K921R, S636N+K627R,S636N, K627R, S636N+Y579W, S636N, Y579W, F638I+Y934G, F638I, Y934G,F638I+K921R, F638I, K921R, F638I+K627R, F638I, K627R, F638I+Y579W,F638I, Y579W, K627R+K51Q, K627R, K51Q, K627R+K451S, K627R, K451S,K627R+A559N, K627R, A559N, K627R+Y579W, K627R, Y579W, Y579W+Y934G,Y579W, Y934G, A651P+F638I, A651P, F638I, P570Q+A651P, P570Q, A651P,P570Q+K921R, P570Q, K921R, P570Q+K627R, P570Q, K627R, P570Q+A559N,P570Q, A559N, P570Q+Y579W, P570Q, Y579W, P570Q+F638I, P570Q, F638I,P570K+Y579W, P570K, Y579W, P570K+F638I, P570K, F638I, P570T+A651P,P570T, A651P, P570T+S636N, P570T, S636N, P570T+Y934G, P570T, Y934G,P570T+F638I, P570T, F638I, P570T+K921R, P570T, K921R, P570T+K627R,P570T, K627R, P570T+A559N, P570T, A559N, P570T+A885F, P570T, A885F,A885F+Y934G, A885F, Y934G, A885F+K627R, A885F, K627R, A559N+Y579W+S636L,A559N, Y579W, S636L, A559N+Y579W+F638I, A559N, Y579W, F638I,A559N+Y579W+D870M, D870M, A559N+Y579W+S560P, S560P, A559N+Y579W+A564I,A564I, A559N+Y579W+P570N, P570N, A559N+Y579W+P570K, P570K,A559N+Y579W+P570R, P570R, A559N+Y579W+P570A, P570A, A559N+Y579W+P570T,P570T, A559N+Y579W+P570S, P570S, A559N+Y579W+P570Q, P570Q,A559N+Y579W+P570H, P570H, and N558E, A559P, A559N, A559H, T561P, A564E,P570A, P570Q, P570R, P570S, P570K, P570T, P570N, Y579W, Y579F, T581M,S616D, K627R, K627M, K627Q, S636N, S636Q, S636R, S636K, S636M, S636H,F638I, F638L, N643D, A651P, A651S, A885F, A885Q, K921R, Y934R, Y934G,N966C, L1011A, K1031I, and A559N+P570A+Y579W, A559N+P570H+Y579W,A559N+P570K+Y579W, A559N+P570N+Y579W, A559N+P570Q+Y579W,A559N+P570R+Y579W, A559N+P570S+Y579W, A559N+P570T+Y579W,A559N+S560P+Y579W, A559N+Y579W+A651P, A559N+Y579W+A651P+Y934G,A559N+Y579W+F638I, A559N+Y579W+K921R, A559N+Y579W+S616D+K921R,A559N+Y579W+S636N, A559N+Y579F, A559N+Y579W, A559N+Y579W+K921R,A559N+Y579W+S616D, F638I+Y934G, K627R+S636N, K627R+Y934G, P570K+Y579W,Q416D+A559N+Y579W+S636N, Q416D, S128X+A559N+Y579W+K627R, S128X,S128X+A559N+Y579W+S636N, Y579W+S636N, V4T, S17A, N18G, F20P, F20N, F20G,F20Y, K51Q, K51H, E53Y, E53P, E53G, Y55M, Y55D, V56M, Y60F, S63F, A71E,579W, T87R, T92S, A120P, N129D, F137L, H182Y, A186P, N189K, K192N,N216D, N216Q, N216R, L226K, G230H, L233H, D247N, G279E, K281R, A283D,N285D, N285G, Q289E, T292A, T292F, T292Y, A294V, Q298E, I302D, I302H,I302V, I302M, H311N, S313D, A346D, A386P, I387T, K388R, K390Q, I403Y,E408D, E408N, E408S, E408P, E408A, E408G, P410G, Q416S, Q416D, N441G,A448E, A448W, A448S, K451S, K451Q, G471S, S472Y, D476R, Q489P, K507R,K512P, S515V, S538C, L555Q, G557R, N558E, A559N, A559P, A559H, A559D,S560P, S560G, T561P, A564E, A564I, V567P, K568R, P570R, P570Q, P570K,P570A, P570T, P570G, P570S, P570H, P570N, I575V, Y579W, Y579F, T581M,S593N, S593E, S595L, S598Q, A599S, I602T, I602D, V603P, S605T, S607C,G609E, S616G, S616D, K627R, K627M, K627Q, K631R, K631A, D635A, D635E,D635M, D635N, D635L, D635W, S636N, S636K, S636L, S636Q, S636R, S636M,S636H, F638N, F638I, F638L, F638V, F638H, F638M, T639G, T639I, T639M,T639Y, T639W, T639P, T639E, T640S, S642N, S642T, N643D, N643H, A651P,A651S, D676H, Q683E, A688G, Y690F, T694A, T697G, R698W, T699A, T706Q,T711S, T711V, T711Y, K713R, W719R, K720H, K744H, K744Q, A749T, K754R,V756Y, V756H, S760G, T781M, N786K, T797S, S810Q, A824D, T825G, N828D,N833D, Q834E, S835A, S835D, V837I, N848D, A868E, A869V, D870V, T872G,T872H, T872W, T872Q, R880K, V881Q, V881T, T883R, T883V, T883C, T883K,Y884H, A885N, A885Q, A885F, T887K, T887S, L888M, V890R, T892P, T892V,R898Q, N905D, F906A, Q912V, N920P, K921R, A924D, V926F, V926P, K927R,S928D, T932A, N933S, N933V, Y934G, Y934R, Y934Q, A937E, V938I, K939V,N941S, A942P, G946R, K948R, Q956Y, Q956S, A957L, A957P, N966C, T972K,M980I, G994D, T999R, L1011A, K1031I, A1037E, S1038G, G1041R, Y1042N, andF1048W.

In some aspects, the present disclosure relates to a detergentcomposition comprising an endoglucanase variant having activity onxanthan gum pre-treated with xanthan lyase; preferably said activitycomprises endoglucanase EC 3.2.1.4 activity, further preferably saidactivity is endoglucanase EC 3.2.1.4 activity.

In some aspects, the present disclosure relates to a detergentcomposition comprising an endoglucanase variant having an improvedstability in a detergent composition compared to a parent endoglucanase(e.g., with SEQ ID NO: 2).

In some aspects, the present disclosure relates to a detergentcomposition comprising an endoglucanase variant having a half-life whichis improved over the parent endoglucanase or a wild-type endoglucanase.

In one embodiment, the endoglucanase variant has a half-life of at leastabout 1.5 hours when measured at a temperature of about 25° C. and in adetergent concentration of about 90%. In a particular embodiment, thehalf-life is measured as described in Example 4. In some aspects, thepresent disclosure relates to a detergent composition comprising anendoglucanase variant having a half-life improvement factor (HIF)of >about 1.0.

In some aspects, the disclosure relates to a detergent compositioncomprising an isolated GH9 endoglucanase variant having activity onxanthan gum pretreated with xanthan lyase. In a further aspect, thecomposition further comprises an isolated polypeptide having xanthanlyase activity.

In some aspects, the present disclosure relates to a detergentcomposition comprising one or more detergent components for degradingxanthan gum.

In some aspects, the present disclosure relates to use of a compositionof the present disclosure, wherein said use is selected from the groupof: use for degrading xanthan gum and use in a cleaning process, such aslaundry or hard surface cleaning such as dish wash.

In some aspects, the present disclosure further relates to the use of adetergent composition of the disclosure for degrading xanthan gum, forwashing or cleaning textiles and/or hard surfaces, such as dish wash,wherein the composition has an enzyme detergency benefit.

In some aspects, the present disclosure also relates to methods ofdegrading xanthan gum using detergent compositions of the presentdisclosure, wherein xanthan gum is on the surface of a hard surface ortextile.

OVERVIEW OF SEQUENCE LISTING

SEQ ID NO: 1 is the DNA sequence of the parent mature endoglucanase froma strain of a Paenibacillus sp.

SEQ ID NO: 2 is the amino acid sequence of mature polypeptide encoded bySEQ ID NO: 1.

SEQ ID NO: 3 is the DNA sequence of the alpha-amylase secretion signalfrom Bacillus licheniformis.

SEQ ID NO: 4 is the amino acid sequence of the alpha-amylase secretionsignal from Bacillus Licheniformis.

Definitions

cDNA: The term “cDNA” means a DNA molecule that can be prepared byreverse transcription from a mature, spliced, mRNA molecule obtainedfrom a eukaryotic or prokaryotic cell. cDNA lacks intron sequences thatmay be present in the corresponding genomic DNA. The initial, primaryRNA transcript is a precursor to mRNA that is processed through a seriesof steps, including splicing, before appearing as mature spliced mRNA.

Cleaning or Detergent Application: the term “cleaning or detergentapplication” means applying the endoglucanase of the application in anycomposition for the purpose of cleaning or washing, by hand, machine orautomated, a hard surface or a textile.

Cleaning Composition: the term “cleaning composition” refers tocompositions that find use in the removal of undesired compounds fromitems to be cleaned, such as textiles, dishes, and hard surfaces. Theterms encompass any materials/compounds selected for the particular typeof cleaning composition desired and the form of the product (e.g.,liquid, gel, powder, granulate, paste, or spray compositions) andincludes, but is not limited to, detergent compositions (e.g., liquidand/or solid laundry detergents and fine fabric detergents; hard surfacecleaning formulations, such as for glass, wood, ceramic and metalcounter tops and windows; carpet cleaners; oven cleaners; fabricfresheners; fabric softeners; and textile and laundry pre-spotters, aswell as dish wash detergents). In addition to the endoglucanase, thedetergent formulation may contain one or more additional enzymes (suchas xanthan lyases, proteases, amylases, lipases, cutinases, cellulases,endoglucanases, xyloglucanases, pectinases, pectin lyases, xanthanases,peroxidaes, haloperoxygenases, catalases and mannanases, or any mixturethereof), and/or components such as surfactants, builders, chelators orchelating agents, bleach system or bleach components, polymers, fabricconditioners, foam boosters, suds suppressors, dyes, perfume, tannishinhibitors, optical brighteners, bactericides, fungicides, soilsuspending agents, anti corrosion agents, enzyme inhibitors orstabilizers, enzyme activators, transferase(s), hydrolytic enzymes,oxido reductases, bluing agents and fluorescent dyes, antioxidants, andsolubilizers.

Coding sequence: The term “coding sequence” means a polynucleotide,which directly specifies the amino acid sequence of a polypeptide. Theboundaries of the coding sequence are generally determined by an openreading frame, which begins with a start codon such as ATG, GTG, or TTGand ends with a stop codon such as TAA, TAG, or TGA. The coding sequencemay be a genomic DNA, cDNA, synthetic DNA, or a combination thereof.

Colour clarification: During washing and wearing loose or broken fiberscan accumulate on the surface of the fabrics. One consequence can bethat the colours of the fabric appear less bright or less intensebecause of the surface contaminations. Removal of the loose or brokenfibers from the textile will partly restore the original colours andlooks of the textile. By the term “colour clarification”, as usedherein, is meant the partial restoration of the initial colours oftextile.

Control sequences: The term “control sequences” means nucleic acidsequences necessary for expression of a polynucleotide encoding a maturepolypeptide of the present disclosure. Each control sequence may benative (i.e., from the same gene) or foreign (i.e., from a differentgene) to the polynucleotide encoding the polypeptide or native orforeign to each other. Such control sequences include, but are notlimited to, a leader, polyadenylation sequence, propeptide sequence,promoter, signal peptide sequence, and transcription terminator. At aminimum, the control sequences include a promoter, and transcriptionaland translational stop signals. The control sequences may be providedwith linkers for the purpose of introducing specific restriction sitesfacilitating ligation of the control sequences with the coding region ofthe polynucleotide encoding a polypeptide.

Corresponding to: The term “corresponding to” as used herein, refers toa way of determining the specific amino acid of a sequence whereinreference is made to a specific amino acid sequence. E.g. for thepurposes of the present disclosure, when references are made to specificamino acid positions, the skilled person would be able to align anotheramino acid sequence to said amino acid sequence that reference has beenmade to, in order to determine which specific amino acid may be ofinterest in said another amino acid sequence. Alignment of another aminoacid sequence with e.g. the sequence as set forth in SEQ ID NO: 2, orany other amino acid sequence listed herein, has been describedelsewhere herein. Alternative alignment methods may be used, and arewell-known for the skilled person.

Degrading xanthan gum and xanthan gum degrading activity: The terms“degrading xanthan gum” and “xanthan gum degrading activity” are usedinterchangebly and are defined as the depolymerization, degradation orbreaking down of xanthan gum into smaller components. The degradation ofxanthan gum can either be the removal of one or more side chainsaccharides, the cutting of the backbone of xanthan gum into smallercomponents or the removal of one or more side chain saccharides and thecutting of the backbone of xanthan gum into smaller components. Apreferred assay for measuring degradation of xanthan gum is the reducingsugar assay as described in example 4 herein. Non-limiting examples ofthe xanthan gum degrading activity include endoglucanase EC 3.2.1.4activity.

Detergent component: the term “detergent component” is defined herein tomean the types of chemicals which can be used in detergent compositions.Examples of detergent components are surfactants, hydrotropes, builders,co-builders, chelators or chelating agents, bleaching system or bleachcomponents, polymers, fabric hueing agents, fabric conditioners, foamboosters, suds suppressors, dispersants, dye transfer inhibitors,fluorescent whitening agents, perfume, optical brighteners,bactericides, fungicides, soil suspending agents, soil release polymers,anti-redeposition agents, enzyme inhibitors or stabilizers, enzymeactivators, antioxidants, and solubilizers. The detergent compositionmay comprise of one or more of any type of detergent component.

Detergent composition: the term “detergent composition” refers tocompositions that find use in the removal of undesired compounds fromitems to be cleaned, such as textiles, dishes, and hard surfaces. Thedetergent composition may be used to e.g. clean textiles, dishes andhard surfaces for both household cleaning and industrial cleaning. Theterms encompass any materials/compounds selected for the particular typeof cleaning composition desired and the form of the product (e.g.,liquid, gel, powder, granulate, paste, or spray compositions) andincludes, but is not limited to, detergent compositions (e.g., liquidand/or solid laundry detergents and fine fabric detergents; hard surfacecleaning formulations, such as for glass, wood, ceramic and metalcounter tops and windows; carpet cleaners; oven cleaners; fabricfresheners; fabric softeners; and textile and laundry pre-spotters, aswell as dish wash detergents). In addition to containing a GH9endoglucanase as described herein and/or xanthan lyase, the detergentformulation may contain one or more additional enzymes (such as xanthanlyases, proteases, amylases, lipases, cutinases, cellulases,endoglucanases, xyloglucanases, pectinases, pectin lyases, xanthanases,peroxidaes, haloperoxygenases, catalases and mannanases, or any mixturethereof), and/or components such as surfactants, builders, chelators orchelating agents, bleach system or bleach components, polymers, fabricconditioners, foam boosters, suds suppressors, dyes, perfume, tannishinhibitors, optical brighteners, bactericides, fungicides, soilsuspending agents, anti corrosion agents, enzyme inhibitors orstabilizers, enzyme activators, transferase(s), hydrolytic enzymes,oxido reductases, bluing agents and fluorescent dyes, antioxidants, andsolubilizers.

Dish wash: The term “dish wash” refers to all forms of washing dishes,e.g. by hand or automatic dish wash. Washing dishes includes, but is notlimited to, the cleaning of all forms of crockery such as plates, cups,glasses, bowls, all forms of cutlery such as spoons, knives, forks andserving utensils as well as ceramics, plastics, metals, china, glass andacrylics.

Dish washing composition: The term “dish washing composition” refers toall forms of compositions for cleaning hard surfaces. The presentdisclosure is not restricted to any particular type of dish washcomposition or any particular detergent.

Endoglucanase: The term “endoglucanase” or “EG” means anendo-1,4-(1,3;1,4)-beta-D-glucan 4-glucanohydrolase (EC 3.2.1.4) thatcatalyzes endohydrolysis of 1,4-beta-D-glycosidic linkages in cellulose,cellulose derivatives (such as carboxymethyl cellulose and hydroxyethylcellulose), lichenin, beta-1,4 bonds in mixed beta-1,3 glucans such ascereal beta-D-glucans, xyloglucans, xanthans and other plant materialcontaining cellulosic components. Endoglucanase activity can bedetermined by measuring reduction in substrate viscosity or increase inreducing ends determined by a reducing sugar assay (Zhang et al., 2006,Biotechnology Advances 24: 452-481). A preferred assay for measuringendoglucanase activity is the reducing sugar assay as described inexample 4 herein. Non-limiting examples of endoglucanases include themature parent endoglucanase having SEQ ID NO: 2.

Enzyme detergency benefit: The term “enzyme detergency benefit” isdefined herein as the advantageous effect an enzyme may add to adetergent compared to the same detergent without the enzyme. Importantdetergency benefits which can be provided by enzymes are stain removalwith no or very little visible soils after washing and or cleaning,prevention or reduction of redeposition of soils released in the washingprocess an effect that also is termed anti-redeposition, restoring fullyor partly the whiteness of textiles, which originally were white butafter repeated use and wash have obtained a greyish or yellowishappearance an effect that also is termed whitening. Textile carebenefits, which are not directly related to catalytic stain removal orprevention of redeposition of soils are also important for enzymedetergency benefits. Examples of such textile care benefits areprevention or reduction of dye transfer from one fabric to anotherfabric or another part of the same fabric an effect that is also termeddye transfer inhibition or anti-backstaining, removal of protruding orbroken fibers from a fabric surface to decrease pilling tendencies orremove already existing pills or fuzz an effect that also is termedanti-pilling, improvement of the fabric-softness, colour clarificationof the fabric and removal of particulate soils which are trapped in thefibers of the fabric or garment. Enzymatic bleaching is a further enzymedetergency benefit where the catalytic activity generally is used tocatalyze the formation of bleaching component such as hydrogen peroxideor other peroxides.

Expression: The term “expression” includes any step involved in theproduction of a polypeptide including, but not limited to,transcription, post-transcriptional modification, translation,post-translational modification, and secretion.

Expression vector: The term “expression vector” means a linear orcircular DNA molecule that comprises a polynucleotide encoding apolypeptide and is operably linked to control sequences that provide forits expression.

Fragment: The term “fragment” means a polypeptide having one or more(e.g., several) amino acids absent from the amino and/or carboxylterminus of a mature polypeptide; wherein the fragment has endoglucanaseactivity. In one aspect, a fragment contains at least about 85%, about86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%,about 93%, about 94% or about 95% of the number of amino acids of themature polypeptide.

Endoglucanase variant having activity on xanthan gum pretreated withxanthan lyase: The term “Endoglucanase variant having activity onxanthan gum pretreated with xanthan lyase” or an “endoglucanase havingactivity on xanthan gum pretreated with xanthan lyase and belonging tothe GH9 class of glycosyl hydrioases” is defined as a polypeptidecomprising a domain belonging to the GH9 class of glycosyl hydrolases,and having activity (e.g. enzymatic activity, xanthan degradingactivity, endoglucanase EC 3.2.1.4 activity) on xanthan gum pretreatedwith xanthan lyase. A preferred assay for measuring activity on xanthangum pretreated with xanthan lyase is disclosed in example 4 herein.

Xanthan lyase variant having activity on xanthan gum: The term “Xanthanlyase variant having activity on xanthan gum” is defined as apolypeptide that cleaves the β-D-mannosyl-β-D-1,4-glucuronosyl bond ofxanthan (e.g., xanthan lyase EC 4.2.2.12 activity). Examples of thexanthan lyase variants having activity on xanthan gum, are xanthan lyasepolypeptides as such. Thus, polypeptides that that cleaves theβ-D-mannosyl-β-D-1,4-glucuronosyl bond of xanthan.

Half-life: the term “half-life” is the time it takes for an enzyme tolose half of its enzymatic activity under a given set of conditions. Itis denoted as T½ and is measured in hours (hrs). Half-lifes can becalculated at a given detegent concentration and storage temperature fora Wild-type control and/or variants, as the degradation follows anexponential decay and the incubation time (hours) is known, i.e.,according to the following formulas:T½(variant)=(Ln(0.5)/Ln(RA-variant/100))*TimeT½(wild-type)=(Ln(0.5)/Ln(RA-wild-type/100))*TimeWherein ‘RA’ is the residual activity in percent and ‘Time’ is theincubation time in hours.

Half-life improvement factor: the term “Half-life improvement factor” or“HIF” is the improvement of half-life of a variant compared to theparent polypeptide, such as the parent endoglucanase. A half-lifeimprovement factor (HIF) under a given set of storage conditions(detergent concentration and temperature) can be calculated as:

${HIF} = \frac{{T\;{1/2}},{variant}}{{T\;{1/2}},{w\; t}}$where the wild-type (wt) is incubated under the same storage condition(detergent concentration and incubation temperature) as the variant. Inthe cases where the difference in stability between wild-type andvariant is too big to accurately assess half-life for both wild-type andvariant using the same incubation time, the incubation time forwild-type and variant is different e.g. about 1 h for wild-type andabout 720 h for the most stable variants. A preferred way of calculatingHIF is also described in example 4 herein.

Hard surface cleaning: The term “Hard surface cleaning” is definedherein as cleaning of hard surfaces wherein hard surfaces may includefloors, tables, walls, roofs etc. as well as surfaces of hard objectssuch as cars (car wash) and dishes (dish wash). Dish washing includesbut are not limited to cleaning of plates, cups, glasses, bowls, andcutlery such as spoons, knives, forks, serving utensils, ceramics,plastics, metals, china, glass and acrylics.

Host cell: The term “host cell” means any cell type that is susceptibleto transformation, transfection, transduction, or the like with anucleic acid construct or expression vector comprising a polynucleotideof the present disclosure. The term “host cell” encompasses any progenyof a parent cell that is not identical to the parent cell due tomutations that occur during replication.

Improved property: The term “improved property” means a characteristicassociated with a variant that is improved compared to the parent. Suchimproved properties include, but are not limited to, catalyticefficiency, catalytic rate, chemical stability, oxidation stability, pHactivity, pH stability, specific activity, stability under storageconditions, chelator stability, substrate binding, substrate cleavage,substrate specificity, substrate stability, surface properties, thermalactivity, and thermostability.

Improved wash performance: The term “improved wash performance” isdefined herein as a (variant) enzyme (also a blend of enzymes, notnecessarily only variants but also backbones, and in combination withcertain cleaning composition etc.) displaying an alteration of the washperformance of a protease variant relative to the wash performance ofthe parent protease variant e.g. by increased stain removal. The term“wash performance” includes wash performance in laundry but also e.g. indish wash.

Isolated: The term “isolated” means a substance in a form or environmentthat does not occur in nature. Non-limiting examples of isolatedsubstances include (1) any non-naturally occurring substance, (2) anysubstance including, but not limited to, any enzyme, variant, nucleicacid, protein, peptide or cofactor, that is at least partially removedfrom one or more or all of the naturally occurring constituents withwhich it is associated in nature; (3) any substance modified by the handof man relative to that substance found in nature; or (4) any substancemodified by increasing the amount of the substance relative to othercomponents with which it is naturally associated (e.g., multiple copiesof a gene encoding the substance; use of a stronger promoter than thepromoter naturally associated with the gene encoding the substance). Anisolated substance may be present in a fermentation broth sample.

Laundering: The term “laundering” relates to both household launderingand industrial laundering and means the process of treating textileswith a solution containing a cleaning or detergent composition of thepresent disclosure. The laundering process can for example be carriedout using e.g. a household or an industrial washing machine or can becarried out by hand.

Mature polypeptide: The term “mature polypeptide” means a polypeptide inits final form following translation and any post-translationalmodifications, such as N-terminal processing, C-terminal truncation,glycosylation, phosphorylation, etc. In one aspect, the maturepolypeptide is amino acids 1 to 1055 of SEQ ID NO: 2.

It is known in the art that a host cell may produce a mixture of two ofmore different mature polypeptides (i.e., with a different C-terminaland/or N-terminal amino acid) expressed by the same polynucleotide. Itis also known in the art that different host cells process polypeptidesdifferently, and thus, one host cell expressing a polynucleotide mayproduce a different mature polypeptide (e.g., having a differentC-terminal and/or N-terminal amino acid) as compared to another hostcell expressing the same polynucleotide.

Mature polypeptide coding sequence: The term “mature polypeptide codingsequence” means a polynucleotide that encodes a mature polypeptidehaving enzymatic activity such as activity on xanthan gum pretreatedwith xanthan lyase or xanthan lyase activity. In one aspect, the maturepolypeptide coding sequence is nucleotides 1 to 3165 of SEQ ID NO: 1.

Mutant: The term “mutant” means a polynucleotide encoding a variant.

Nucleic acid construct: The term “nucleic acid construct” means anucleic acid molecule, either single- or double-stranded, which isisolated from a naturally occurring gene or is modified to containsegments of nucleic acids in a manner that would not otherwise exist innature or which is synthetic, which comprises one or more controlsequences.

Operably linked: The term “operably linked” means a configuration inwhich a control sequence is placed at an appropriate position relativeto the coding sequence of a polynucleotide such that the controlsequence directs expression of the coding sequence.

Parent: The term “parent” or “parent endoglucanase” means anypolypeptide with endoglucanase activity to which an alteration is madeto produce the enzyme variants of the present disclosure. In one aspect,the parent is an endoglucanase having the identical amino acid sequenceof the variant, but not having the alterations at one or more of thespecified positions. It will be understood, that the expression “havingidentical amino acid sequence” relates to about 100% sequence identity.Non-limiting examples of parent endoglucanases include the mature parentendoglucanase having SEQ ID NO: 2.

Sequence identity: The relatedness between two amino acid sequences orbetween two nucleotide sequences is described by the parameter “sequenceidentity”. For purposes of the present disclosure, the sequence identitybetween two amino acid sequences is determined using theNeedleman-Wunsch algorithm (Needleman and Wunsch, 1970, J. Mol. Biol.48: 443-453) as implemented in the Needle program of the EMBOSS package(EMBOSS: The European Molecular Biology Open Software Suite, Rice etal., 2000, Trends Genet. 16: 276-277), preferably version 5.0.0 orlater. The parameters used are gap open penalty of 10, gap extensionpenalty of 0.5, and the EBLOSUM62 (EMBOSS version of BLOSUM62)substitution matrix. The output of Needle labeled “longest identity”(obtained using the—nobrief option) is used as the percent identity andis calculated as follows:(Identical Residues×100)/(Length of Alignment−Total Number of Gaps inAlignment)

For purposes of the present disclosure, the sequence identity betweentwo deoxyribonucleotide sequences is determined using theNeedleman-Wunsch algorithm (Needleman and Wunsch, 1970, supra) asimplemented in the Needle program of the EMBOSS package (EMBOSS: TheEuropean Molecular Biology Open Software Suite, Rice et al., 2000,supra), preferably version 5.0.0 or later. The parameters used are gapopen penalty of 10, gap extension penalty of 0.5, and the EDNAFULL(EMBOSS version of NCBI NUC4.4) substitution matrix. The output ofNeedle labeled “longest identity” (obtained using the—nobrief option) isused as the percent identity and is calculated as follows:(Identical Deoxyribonucleotides×100)/(Length of Alignment−Total Numberof Gaps in Alignment)

Stringency conditions: The different strigency conditions are defined asfollows.

The term “very low stringency conditions” means for probes of at leastabout 100 nucleotides in length, prehybridization and hybridization atabout 42° C. in 5×SSPE, about 0.3% SDS, 200 micrograms/ml sheared anddenatured salmon sperm DNA, and about 25% formamide, following standardSouthern blotting procedures for from about 12 to about 24 hours. Thecarrier material is finally washed three times each for about 15 minutesusing 2×SSC, about 0.2% SDS at about 45° C.

The term “low stringency conditions” means for probes of at least about100 nucleotides in length, prehybridization and hybridization at about42° C. in 5×SSPE, about 0.3% SDS, about 200 micrograms/ml sheared anddenatured salmon sperm DNA, and about 25% formamide, following standardSouthern blotting procedures for from about 12 to about 24 hours. Thecarrier material is finally washed three times each for about 15 minutesusing 2×SSC, about 0.2% SDS at about 50° C.

The term “medium stringency conditions” means for probes of at leastabout 100 nucleotides in length, prehybridization and hybridization atabout 42° C. in 5×SSPE, 0.3% SDS, 200 micrograms/ml sheared anddenatured salmon sperm DNA, and about 35% formamide, following standardSouthern blotting procedures for from about 12 to about 24 hours. Thecarrier material is finally washed three times each for about 15 minutesusing 2×SSC, about 0.2% SDS at about 55° C.

The term “medium-high stringency conditions” means for probes of atleast about 100 nucleotides in length, prehybridization andhybridization at about 42° C. in 5×SSPE, about 0.3% SDS, about 200micrograms/ml sheared and denatured salmon sperm DNA, and about 35%formamide, following standard Southern blotting procedures for fromabout 12 to about 24 hours. The carrier material is finally washed threetimes each for about 15 minutes using 2×SSC, about 0.2% SDS at about 60°C.

The term “high stringency conditions” means for probes of at least about100 nucleotides in length, prehybridization and hybridization at about42° C. in 5×SSPE, about 0.3% SDS, 200 micrograms/ml sheared anddenatured salmon sperm DNA, and about 50% formamide, following standardSouthern blotting procedures for from about 12 to about 24 hours. Thecarrier material is finally washed three times each for about 15 minutesusing 2×SSC, about 0.2% SDS at about 65° C.

The term “very high stringency conditions” means for probes of at leastabout 100 nucleotides in length, prehybridization and hybridization atabout 42° C. in 5×SSPE, about 0.3% SDS, about 200 micrograms/ml shearedand denatured salmon sperm DNA, and about 50% formamide, followingstandard Southern blotting procedures for from about 12 to about 24hours. The carrier material is finally washed three times each for about15 minutes using 2×SSC, about 0.2% SDS at about 70° C.

Subsequence: The term “subsequence” means a polynucleotide having one ormore (e.g., several) nucleotides absent from the 5′ and/or 3′ end of amature polypeptide coding sequence; wherein the subsequence encodes afragment having enzymatic activity, such as activity on xanthan gumpretreated with xanthan lyase or xanthan lyase activity.

Textile: The term “textile” means any textile material including yarns,yarn intermediates, fibers, non-woven materials, natural materials,synthetic materials, and any other textile material, fabrics made ofthese materials and products made from fabrics (e.g., garments and otherarticles). The textile or fabric may be in the form of knits, wovens,denims, non-wovens, felts, yarns, and towelling. The textile may becellulose based such as natural cellulosics, including cotton,flax/linen, jute, ramie, sisal or coir or manmade cellulosics (e.g.originating from wood pulp) including viscose/rayon, ramie, celluloseacetate fibers (tricell), lyocell or blends thereof. The textile orfabric may also be non-cellulose based such as natural polyamidesincluding wool, camel, cashmere, mohair, rabit and silk or syntheticpolymer such as nylon, aramid, polyester, acrylic, polypropylen andspandex/elastane, or blends thereof as well as blend of cellulose basedand non-cellulose based fibers. Examples of blends are blends of cottonand/or rayon/viscose with one or more companion material such as wool,synthetic fibers (e.g. polyamide fibers, acrylic fibers, polyesterfibers, polyvinyl alcohol fibers, polyvinyl chloride fibers,polyurethane fibers, polyurea fibers, aramid fibers), andcellulose-containing fibers (e.g. rayon/viscose, ramie, flax/linen,jute, cellulose acetate fibers, lyocell). Fabric may be conventionalwashable laundry, for example stained household laundry. When the termfabric or garment is used it is intended to include the broader termtextiles as well.

Textile care benefit: “Textile care benefits”, which are not directlyrelated to catalytic stain removal or prevention of redeposition ofsoils, are also important for enzyme detergency benefits. Examples ofsuch textile care benefits are prevention or reduction of dye transferfrom one textile to another textile or another part of the same textilean effect that is also termed dye transfer inhibition oranti-backstaining, removal of protruding or broken fibers from a textilesurface to decrease pilling tendencies or remove already existing pillsor fuzz an effect that also is termed anti-pilling, improvement of thetextile-softness, colour clarification of the textile and removal ofparticulate soils which are trapped in the fibers of the textile.Enzymatic bleaching is a further enzyme detergency benefit where thecatalytic activity generally is used to catalyze the formation ofbleaching component such as hydrogen peroxide or other peroxides orother bleaching species.

Variant: The term “variant” means a polypeptide (e.g., a GH9endoglucanase polypeptide) comprising an alteration i.e., asubstitution, insertion, and/or deletion, at one or more (e.g., several)positions. A substitution means replacement of the amino acid occupyinga position with a different amino acid; a deletion means removal of theamino acid occupying a position; and an insertion means adding one ormore (e.g. several) amino acids e.g., 1-5 amino acids adjacent to andimmediately following the amino acid occupying a position. Non-limitingexamples of endoglucanase variants described herein includeendoglucanase variants having an activity on xanthan gum pretreated withxanthan lyase. Non-limiting examples of variants described hereinfurther include variants having at least about 20%, e.g., at least about40%, at least about 50%, at least about 60%, at least about 70%, atleast about 80%, at least about 90%, at least about 95%, or at leastabout 100% endoglucanase activity of the mature parent having SEQ ID NO:2. A preferred assay for measuring activity on xanthan gum pretreatedwith xanthan lyase is disclosed in example 4 herein.

Stability: The term “stability” means resistance or the degree ofresistance to change, unfolding, disintegration, denaturation oractivity loss. Non-limiting examples of stability include conformationalstability, storage stability and stability during use, e.g. during awash process and reflects the stability of a polypeptide (e.g. anendoglucanase variant as contemplated herein) as a function of time,e.g. how much activity is retained when said polypeptide (e.g. saidendoglucanase variant) is kept in solution, in particular in a detergentsolution. The stability is influenced by many factors, e.g. presence ofchelator(s), pH, temperature, detergent composition, e.g. amount ofbuilders, surfactants, chelators etc. The endoglucanase stability may bemeasured using a half-life improvement factor (HIF) as described inexample 4 herein. The endoglucanase stability may also be measured usinga reducing sugar assay as described in example 4 herein.

Improved stability: The term “improved stability” or “increasedstability” is defined herein as increased stability in a detergentcomposition (e.g., in solutions, e.g. in the presence of a chelator,e.g. EDTA or citrate), relative to the stability of the parentendoglucanase, relative to an endoglucanase having the identical aminoacid sequence of the variant, but not having the alterations at one ormore of the specified positions, or relative to SEQ ID NO: 2. The terms“improved stability” and “increased stability” includes “improvedchemical stability”, “detergent stability” and “improved detergentstability.

Improved chemical stability: The term “improved chemical stability” isdefined herein as a variant enzyme displaying retention of enzymaticactivity after a period of incubation in the presence of a chemical orchemicals, either naturally occurring or synthetic, which reduces theenzymatic activity of the parent enzyme. Improved chemical stability mayalso result in variants being more able (e.g., better that the parent)to catalyze a reaction in the presence of such chemicals. In aparticular aspect of the disclosure the improved chemical stability isan improved stability in a detergent, in particular in a liquiddetergent. The term “detergent stability” or “improved detergentstability is in particular an improved stability of the endoglucanasecompared to the parent endoglucanase, when an endoglucanase variant ofthe present disclosure is mixed into a liquid detergent formulation,especially into a liquid detergent formulation comprising a chelator(e.g. EDTA or citrate).

Conformational stability: The term “conformational stability” meansresistance or a degree of resistance to conformational change, unfoldingor disintegration. Accordingly, the term “less conformationally stable”means less resistant or having lesser degree of resistance toconformational change, unfolding or disintegration.

Instability: The term “instability” means lack of stability.Non-limiting examples of instability include conformational instability,unfolding, denaturation, desintegration, activity loss.

Chelator-induced instability region: The term “chelator-inducedinstability region” means any region of a polypeptide contibuting toinstability of said polypeptide in the presence of a chelator.Non-limiting examples of chelators include EDTA(Ethylenediaminetetraacetic acid) and citrate. Non-limiting examples ofchelator-induced instability regions include any region of a polypeptidehaving one or more of the following features: in the presence of achelator it is less conformationally stable than one or more or all ofits adjacent regions; and/or in the presence of a chelator it is moreexposed to said chelator than one or more or all of its adjacentregions; and/or in the presence of a chelator it is more accessible tosaid chelator than one or more or all of its adjacent regions; and/or inthe presence of a chelator it is more conformationally dynamic than oneor more or all of its adjacent regions; and/or in the presence of achelator it is more receptive to deuterium incorporation than one ormore or all of its adjacent regions. Non-limiting examples ofchelator-induced instability regions further include any region of apolypeptide responsible for chelator-induced instability. Non-limitingexamples of chelator-induced instability regions of a matureendoglucanase (e.g. having SEQ ID NO: 2) include: region 1 correspondingto amino acids 95 to 105 of SEQ ID NO: 2, region 2 corresponding toamino acids 115 to 138 of SEQ ID NO: 2, region 3 corresponding to aminoacids 210 to 251 of SEQ ID NO: 2, region 4 corresponding to amino acids267 to 301 of SEQ ID NO: 2, region 5 corresponding to amino acids 339 to361 of SEQ ID NO: 2, region 6 corresponding to amino acids 547 to 595 ofSEQ ID NO: 2, region 7 corresponding to amino acids 612 to 660 of SEQ IDNO: 2, region 8 corresponding to amino acids 806 to 828 of SEQ ID NO: 2,and region 9 corresponding to amino acids 839 to 1042 of SEQ ID NO: 2.Non-limiting examples of regions adjacent to chelator-inducedinstability regions of a mature endoglucanase (e.g. having SEQ ID NO: 2)include: region 10 corresponding to amino acids 1 to 94 of SEQ ID NO: 2,region 11 corresponding to amino acids 106 to 114 of SEQ ID NO: 2,region 12 corresponding to amino acids 139 to 209 of SEQ ID NO: 2,region 13 corresponding to amino acids 252 to 266 of SEQ ID NO: 2,region 14 corresponding to amino acids 302 to 338 of SEQ ID NO: 2,region 15 corresponding to amino acids 362 to 546 of SEQ ID NO: 2,region 16 corresponding to amino acids 596 to 611 of SEQ ID NO: 2,region 17 corresponding to amino acids 661 to 805 of SEQ ID NO: 2,region 18 corresponding to amino acids 829 to 838 of SEQ ID NO: 2, andregion 19 corresponding to amino acids 1043 to 1055 of SEQ ID NO: 2.

Adjacent region: The term “adjacent region” means any region of apolypeptide that is not a chelator-induced instability region.Non-limiting examples of adjacent regions of a mature endoglucanase(e.g. having SEQ ID NO: 2) include: region 10 corresponding to aminoacids 1 to 94 of SEQ ID NO: 2, region 11 corresponding to amino acids106 to 114 of SEQ ID NO: 2, region 12 corresponding to amino acids 139to 209 of SEQ ID NO: 2, region 13 corresponding to amino acids 252 to266 of SEQ ID NO: 2, region 14 corresponding to amino acids 302 to 338of SEQ ID NO: 2, region 15 corresponding to amino acids 362 to 546 ofSEQ ID NO: 2, region 16 corresponding to amino acids 596 to 611 of SEQID NO: 2, region 17 corresponding to amino acids 661 to 805 of SEQ IDNO: 2, region 18 corresponding to amino acids 829 to 838 of SEQ ID NO:2, and region 19 corresponding to amino acids 1043 to 1055 of SEQ ID NO:2.

Chelator exposure: The term “chelator exposure” means concentration oramount of a chelator that reaches a polypeptide. Accordingly, in thecontext of the present disclosure the term “more exposed to a chelator”means that chelator exposure of a particular region (e.g. achelator-induced instability region) is greater than a chelator exposureof a different region (e.g. an adjacent region). In one aspect, chelatorexposure can be expressed in numerical terms of concentration, duration,and frequency (e.g. for chemical agents, e.g. chelators) or intensity.

Chelator accessibility: The term “chelator accessibility” encompasesopenness to the influence by a chelator and easiness of approach bychelator. Accordingly, in the context of the present disclosure the term“more accessible to a chelator” means that chelator accessibility of aparticular region (e.g. a chelator-induced instability region) isgreater than a chelator accessibility of a different region (e.g. anadjacent region).

Conformational dynamics: The term “conformational dynamics” encompassesvibrations, structural rearrangements and transitions of a polypeptide(e.g. in solution). Accordingly, in the context of the presentdisclosure the term “more conformationally dynamic” means thatconformational dynamics of a particular region (e.g. a chelator-inducedinstability region) is greater than conformational dynamics of adifferent region (e.g. an adjacent region).

Receptiveness to deuterium incorporation: The term “receptiveness todeuterium incorporation” means amount of hydrogen atoms replaced by adeuterium atoms during hydrogen-deuterium exchange. Said amount can bemeasured in relative (e.g. compared to another amount) or absolute (e.g.expressed numerically) terms. Accordingly, in the context of the presentdisclosure the term “more receptive to deuterium incorporation” meansthat receptiveness to deuterium incorporation of a particular region(e.g. a chelator-induced instability region) is greater thanreceptiveness to deuterium incorporation of a different region (e.g. anadjacent region).

Wash performance: The term “wash performance” is used as an enzyme'sability to remove stains present on the object to be cleaned during e.g.wash or hard surface cleaning.

Conventions for Designation of Variants

For purposes of the present disclosure, the mature polypeptide disclosedin SEQ ID NO: 2 is used to determine the corresponding amino acidresidue in another endoglucanase. The amino acid sequence of anotherendoglucanase is aligned with the mature polypeptide disclosed in SEQ IDNO: 2, and based on the alignment, the amino acid position numbercorresponding to any amino acid residue in the mature polypeptidedisclosed in SEQ ID NO: 2 is determined using the Needleman-Wunschalgorithm (Needleman and Wunsch, 1970, J. Mol. Biol. 48: 443-453) asimplemented in the Needle program of the EMBOSS package (EMBOSS: TheEuropean Molecular Biology Open Software Suite, Rice et al., 2000,Trends Genet. 16: 276-277), preferably version 5.0.0 or later. Theparameters used are gap open penalty of 10, gap extension penalty of0.5, and the EBLOSUM62 (EMBOSS version of BLOSUM62) substitution matrix.

Identification of the corresponding amino acid residue in anotherendoglucanase can be determined by an alignment of multiple polypeptidesequences using several computer programs including, but not limited to,MUSCLE (multiple sequence comparison by log-expectation; version 3.5 orlater; Edgar, 2004, Nucleic Acids Research 32: 1792-1797), MAFFT(version 6.857 or later; Katoh and Kuma, 2002, Nucleic Acids Research30: 3059-3066; Katoh et al., 2005, Nucleic Acids Research 33: 511-518;Katoh and Toh, 2007, Bioinformatics 23: 372-374; Katoh et al., 2009,Methods in Molecular Biology 537: 39-64; Katoh and Toh, 2010,Bioinformatics 26: 1899-1900), and EMBOSS EMMA employing ClustalW (1.83or later; Thompson et al., 1994, Nucleic Acids Research 22: 4673-4680),using their respective default parameters.

When the other enzyme has diverged from the mature polypeptide of SEQ IDNO: 2 such that traditional sequence-based comparison fails to detecttheir relationship (Lindahl and Elofsson, 2000, J. Mol. Biol. 295:613-615), other pairwise sequence comparison algorithms can be used.Greater sensitivity in sequence-based searching can be attained usingsearch programs that utilize probabilistic representations ofpolypeptide families (profiles) to search databases. For example, thePSI-BLAST program generates profiles through an iterative databasesearch process and is capable of detecting remote homologs (Atschul etal., 1997, Nucleic Acids Res. 25: 3389-3402). Even greater sensitivitycan be achieved if the family or superfamily for the polypeptide has oneor more representatives in the protein structure databases. Programssuch as GenTHREADER (Jones, 1999, J. Mol. Biol. 287: 797-815; McGuffinand Jones, 2003, Bioinformatics 19: 874-881) utilize information from avariety of sources (PSI-BLAST, secondary structure prediction,structural alignment profiles, and solvation potentials) as input to aneural network that predicts the structural fold for a query sequence.Similarly, the method of Gough et al., 2000, J. Mol. Biol. 313: 903-919,can be used to align a sequence of unknown structure with thesuperfamily models present in the SCOP database. These alignments can inturn be used to generate homology models for the polypeptide, and suchmodels can be assessed for accuracy using a variety of tools developedfor that purpose.

For proteins of known structure, several tools and resources areavailable for retrieving and generating structural alignments. Forexample the SCOP superfamilies of proteins have been structurallyaligned, and those alignments are accessible and downloadable. Two ormore protein structures can be aligned using a variety of algorithmssuch as the distance alignment matrix (Holm and Sander, 1998, Proteins33: 88-96) or combinatorial extension (Shindyalov and Bourne, 1998,Protein Engineering 11: 739-747), and implementation of these algorithmscan additionally be utilized to query structure databases with astructure of interest in order to discover possible structural homologs(e.g., Holm and Park, 2000, Bioinformatics 16: 566-567).

In describing the variants of the present disclosure, the nomenclaturedescribed below is adapted for ease of reference. The accepted IUPACsingle letter or three letter amino acid abbreviation is employed.

Substitutions.

For an amino acid substitution, the following nomenclature is used:Original amino acid, position, substituted amino acid. Accordingly, thesubstitution of threonine at position 226 with alanine is designated as“Thr226Ala” or “T226A”. Multiple mutations are separated by additionmarks (“+”), e.g., “Gly205Arg+Ser411Phe” or “G205R+S411F”, representingsubstitutions at positions 205 and 411 of glycine (G) with arginine (R)and serine (S) with phenylalanine (F), respectively.

Deletions.

For an amino acid deletion, the following nomenclature is used: Originalamino acid, position, *. Accordingly, the deletion of glycine atposition 195 is designated as “Gly195*” or “G195*”. Multiple deletionsare separated by addition marks (“+”), e.g., “Gly195*+Ser411*” or“G195*+S411*”.

Insertions.

For an amino acid insertion, the following nomenclature is used:Original amino acid, position, original amino acid, inserted amino acid.Accordingly, the insertion of lysine after glycine at position 195 isdesignated “Gly195GlyLys” or “G195GK”. An insertion of multiple aminoacids is designated [Original amino acid, position, original amino acid,inserted amino acid #1, inserted amino acid #2; etc.]. For example, theinsertion of lysine and alanine after glycine at position 195 isindicated as “Gly195GlyLysAla” or “G195GKA”.

In such cases the inserted amino acid residue(s) are numbered by theaddition of lower case letters to the position number of the amino acidresidue preceding the inserted amino acid residue(s). In the aboveexample, the sequence would thus be:

Parent: Variant: 195 195 195a 195b G G-K-A

Multiple Alterations.

Variants comprising multiple alterations are separated by addition marks(“+”), e.g., “Arg170Tyr+Gly195Glu” or “R170Y+G195E” representing asubstitution of arginine and glycine at positions 170 and 195 withtyrosine and glutamic acid, respectively.

Different Alterations.

Where different alterations can be introduced at a position, thedifferent alterations are separated by a comma, e.g., “Arg170Tyr,Glu”represents a substitution of arginine at position 170 with tyrosine orglutamic acid. Thus, “Tyr167Gly,Ala+Arg170Gly,Ala” designates thefollowing variants: “Tyr167Gly+Arg170Gly”, “Tyr167Gly+Arg170Ala”,“Tyr167Ala+Arg170Gly”, and “Tyr167Ala+Arg170Ala”.

The known xanthan endoglucanase having SEQ ID NO: 2 is a large enzyme(>1000 residues), it is therefore extremely laborious and expensive totarget its properties for improvement of, e.g. stability in a detergentcomposition, e.g. in the presence of a chelator. In some aspects, thepresent disclosure narrows down the number of residues to target whentrying to stabilize endoglucanase molecules using protein engineering toa region selected from the group of: region 1 corresponding to aminoacids 95 to 105 of SEQ ID NO: 2, region 2 corresponding to amino acids115 to 138 of SEQ ID NO: 2, region 3 corresponding to amino acids 210 to251 of SEQ ID NO: 2, region 4 corresponding to amino acids 267 to 301 ofSEQ ID NO: 2, region 5 corresponding to amino acids 339 to 361 of SEQ IDNO: 2, region 6 corresponding to amino acids 547 to 595 of SEQ ID NO: 2,region 7 corresponding to amino acids 612 to 660 of SEQ ID NO: 2, region8 corresponding to amino acids 806 to 828 of SEQ ID NO: 2, and region 9corresponding to amino acids 839 to 1042 of SEQ ID NO: 2.

In one embodiment, the present disclosure dramatically narrows down thenumber of residues to target when trying to stabilize endoglucanasemolecules using protein engineering.

In another embodiment, the present disclosure provides detergentcompositions comprising variants of an endoglucanase which hassignificantly improved stability as compared to the parentendoglucanase, such as the wild-type endoglucanase. Such improvedstability may be measured as improved half-life of the variant comparedto the parent endoglucanase, such as a wild-type endoglucanase.Furthermore, the stability of the variant is also proven to be improvedin the presence of a protease, which normally would cleave proteins. Thepresent disclosure discloses variants that have been modified so thatthey have an improved stability towards protease cleavage.

Variants

In one embodiment, chelator-induced instability regions in the proteinsequence of the known xanthan endoglucanase having SEQ ID NO: 2 that areaffected when the molecule is incubated in a buffer with EDTA, are thefollowing: region 1 corresponding to amino acids 95 to 105 of SEQ ID NO:2, region 2 corresponding to amino acids 115 to 138 of SEQ ID NO: 2,region 3 corresponding to amino acids 210 to 251 of SEQ ID NO: 2, region4 corresponding to amino acids 267 to 301 of SEQ ID NO: 2, region 5corresponding to amino acids 339 to 361 of SEQ ID NO: 2, region 6corresponding to amino acids 547 to 595 of SEQ ID NO: 2, region 7corresponding to amino acids 612 to 660 of SEQ ID NO: 2, region 8corresponding to amino acids 806 to 828 of SEQ ID NO: 2, and region 9corresponding to amino acids 839 to 1042 of SEQ ID NO: 2. Thisembodiment relates to an important guidance on where to mutate anendoglucanase in order to stabilize its molecule in a detergent, e.g.detergent composition comprising a chelator, e.g. EDTA or citrate.

Accordingly, in one embodiment the present disclosure relates to adetergent composition comprising an endoglucanase variant, comprising analteration (e.g., a substitution, deletion or insertion) at one or morepositions in a region selected from the group of: region 1 correspondingto amino acids 95 to 105 of SEQ ID NO: 2, region 2 corresponding toamino acids 115 to 138 of SEQ ID NO: 2, region 3 corresponding to aminoacids 210 to 251 of SEQ ID NO: 2, region 4 corresponding to amino acids267 to 301 of SEQ ID NO: 2, region 5 corresponding to amino acids 339 to361 of SEQ ID NO: 2, region 6 corresponding to amino acids 547 to 595 ofSEQ ID NO: 2, region 7 corresponding to amino acids 612 to 660 of SEQ IDNO: 2, region 8 corresponding to amino acids 806 to 828 of SEQ ID NO: 2,and region 9 corresponding to amino acids 839 to 1042 of SEQ ID NO: 2,wherein said variant has at least about 60% and less than about 100%sequence identity to SEQ ID NO: 2; preferably said endoglucanase varianthas activity on xanthan gum pretreated with xanthan lyase, furtherpreferably said activity is a xanthan gum degrading activity.

In one embodiment the present disclosure relates to a detergentcomposition comprising an endoglucanase variant, comprising analteration (e.g., a substitution, deletion or insertion) at one or morepositions in a region selected from the group of:

i) region 1 corresponding to amino acids 95 to 105 of SEQ ID NO: 2,e.g., said alteration at one or more positions selected from the groupof positions: 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, and 105,wherein said positions correspond to amino acid positions of SEQ ID NO:2 (e.g., using the numbering of SEQ ID NO: 2),

ii) region 2 corresponding to amino acids 115 to 138 of SEQ ID NO: 2,e.g., said alteration at one or more positions selected from the groupof positions: 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125,126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, and 138,wherein said positions correspond to amino acid positions of SEQ ID NO:2 (e.g., using the numbering of SEQ ID NO: 2),

iii) region 3 corresponding to amino acids 210 to 251 of SEQ ID NO: 2,e.g., said alteration at one or more positions selected from the groupof positions: 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220,221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234,235, 236, 237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248,249, 250, and 251, wherein said positions correspond to amino acidpositions of SEQ ID NO: 2 (e.g., using the numbering of SEQ ID NO: 2),

iv) region 4 corresponding to amino acids 267 to 301 of SEQ ID NO: 2,e.g., said alteration at one or more positions selected from the groupof positions: 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277,278, 279, 280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291,292, 293, 294, 295, 296, 297, 298, 299, 300, and 301, wherein saidpositions correspond to amino acid positions of SEQ ID NO: 2 (e.g.,using the numbering of SEQ ID NO: 2),

v) region 5 corresponding to amino acids 339 to 361 of SEQ ID NO: 2,e.g., said alteration at one or more positions selected from the groupof positions: 339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349,350, 351, 352, 353, 354, 355, 356, 357, 358, 359, 360, 361, wherein saidpositions correspond to amino acid positions of SEQ ID NO: 2 (e.g.,using the numbering of SEQ ID NO: 2),

vi) region 6 corresponding to amino acids 547 to 595 of SEQ ID NO: 2,e.g., said alteration at one or more positions selected from the groupof positions: 547, 548, 549, 550, 551, 552, 553, 554, 555, 556, 557,558, 559, 560, 561, 562, 563, 564, 565, 566, 567, 568, 569, 570, 571,572, 573, 574, 575, 576, 577, 578, 579, 580, 581, 582, 583, 584, 585,586, 587, 588, 589, 590, 591, 592, 593, 594, and 595, wherein saidpositions correspond to amino acid positions of SEQ ID NO: 2 (e.g.,using the numbering of SEQ ID NO: 2),

vii) region 7 corresponding to amino acids 612 to 660 of SEQ ID NO: 2,e.g., said alteration at one or more positions selected from the groupof positions: 612, 613, 614, 615, 616, 617, 618, 619, 620, 621, 622,623, 624, 625, 626, 627, 628, 629, 630, 631, 632, 633, 634, 635, 636,637, 638, 639, 640, 641, 642, 643, 644, 645, 646, 647, 648, 649, 650,651, 652, 653, 654, 655, 656, 657, 658, 659, and 660, wherein saidpositions correspond to amino acid positions of SEQ ID NO: 2 (e.g.,using the numbering of SEQ ID NO: 2),

viii) region 8 corresponding to amino acids 806 to 828 of SEQ ID NO: 2,e.g., said alteration at one or more positions selected from the groupof positions: 806, 807, 808, 809, 810, 811, 812, 813, 814, 815, 816,817, 818, 819, 820, 821, 822, 823, 824, 825, 826, 827, and 828, whereinsaid positions correspond to amino acid positions of SEQ ID NO: 2 (e.g.,using the numbering of SEQ ID NO: 2), and

ix) region 9 corresponding to amino acids 839 to 1042 of SEQ ID NO: 2,e.g., said alteration at one or more positions selected from the groupof positions: 839, 840, 841, 842, 843, 844, 845, 846, 847, 848, 849,850, 851, 852, 853, 854, 855, 856, 857, 858, 859, 860, 861, 862, 863,864, 865, 866, 867, 868, 869, 870, 871, 872, 873, 874, 875, 876, 877,878, 879, 880, 881, 882, 883, 884, 885, 886, 887, 888, 889, 890, 891,892, 893, 894, 895, 896, 897, 898, 899, 900, 901, 902, 903, 904, 905,906, 907, 908, 909, 910, 911, 912, 913, 914, 915, 916, 917, 918, 919,920, 921, 922, 923, 924, 925, 926, 927, 928, 929, 930, 931, 932, 933,934, 935, 936, 937, 938, 939, 940, 941, 942, 943, 944, 945, 946, 947,948, 949, 950, 951, 952, 953, 954, 955, 956, 957, 958, 959, 960, 961,962, 963, 964, 965, 966, 967, 968, 969, 970, 971, 972, 973, 974, 975,976, 977, 978, 979, 980, 981, 982, 983, 984, 985, 986, 987, 988, 989,990, 991, 992, 993, 994, 995, 996, 997, 998, 999, 1000, 1001, 1002,1003, 1004, 1005, 1006, 1007, 1008, 1009, 1010, 1011, 1012, 1013, 1014,1015, 1016, 1017, 1018, 1019, 1020, 1021, 1022, 1023, 1024, 1025, 1026,1027, 1028, 1029, 1030, 1031, 1032, 1033, 1034, 1035, 1036, 1037, 1038,1039, 1040, 1041, and 1042, wherein said positions correspond to aminoacid positions of SEQ ID NO: 2 (e.g., using the numbering of SEQ ID NO:2).

In one embodiment the present disclosure relates to a detergentcomposition comprising an endoglucanase variant, comprising analteration (e.g., a substitution, deletion or insertion) at one or morepositions in one or more regions selected from the group of: region 1corresponding to amino acids 95 to 105 of SEQ ID NO: 2, region 2corresponding to amino acids 115 to 138 of SEQ ID NO: 2, region 3corresponding to amino acids 210 to 251 of SEQ ID NO: 2, region 4corresponding to amino acids 267 to 301 of SEQ ID NO: 2, region 5corresponding to amino acids 339 to 361 of SEQ ID NO: 2, region 6corresponding to amino acids 547 to 595 of SEQ ID NO: 2, region 7corresponding to amino acids 612 to 660 of SEQ ID NO: 2, region 8corresponding to amino acids 806 to 828 of SEQ ID NO: 2, and region 9corresponding to amino acids 839 to 1042 of SEQ ID NO: 2, wherein saidvariant has at least about 60% and less than about 100% sequenceidentity to SEQ ID NO: 2; preferably said endoglucanase variant hasactivity on xanthan gum pretreated with xanthan lyase, furtherpreferably said activity is a xanthan gum degrading activity.

In one embodiment the present disclosure relates to a detergentcomposition comprising a parent endoglucanase of the disclosure (e.g.,SEQ ID NO: 2) having an alteration (e.g., a substitution, deletion orinsertion) at one or more positions in a region selected from the groupof regions 1-9, wherein said region is a chelator-induced instabilityregion, preferably said chelator-induced instability region has one ormore of the following features: in the presence of a chelator it is lessconformationally stable than one or more or all of its adjacent regions;and/or in the presence of a chelator it is more exposed to said chelatorthan one or more or all of its adjacent regions; and/or in the presenceof a chelator it is more accessible to said chelator than one or more orall of its adjacent regions; and/or in the presence of a chelator it ismore conformationally dynamic than one or more or all of its adjacentregions; and/or in the presence of a chelator it is more receptive todeuterium incorporation than one or more or all of its adjacent regions;further preferably said adjacent region is selected from the group of:region 10 corresponding to amino acids 1 to 94 of SEQ ID NO: 2, region11 corresponding to amino acids 106 to 114 of SEQ ID NO: 2, region 12corresponding to amino acids 139 to 209 of SEQ ID NO: 2, region 13corresponding to amino acids 252 to 266 of SEQ ID NO: 2, region 14corresponding to amino acids 302 to 338 of SEQ ID NO: 2, region 15corresponding to amino acids 362 to 546 of SEQ ID NO: 2, region 16corresponding to amino acids 596 to 611 of SEQ ID NO: 2, region 17corresponding to amino acids 661 to 805 of SEQ ID NO: 2, region 18corresponding to amino acids 829 to 838 of SEQ ID NO: 2, and region 19corresponding to amino acids 1043 to 1055 of SEQ ID NO: 2; further mostpreferably said chelator is EDTA or citrate.

In one embodiment the adjacent regions referred to herein can be one ormore or all of the following: region 10 corresponding to amino acids 1to 94 of SEQ ID NO: 2, region 11 corresponding to amino acids 106 to 114of SEQ ID NO: 2, region 12 corresponding to amino acids 139 to 209 ofSEQ ID NO: 2, region 13 corresponding to amino acids 252 to 266 of SEQID NO: 2, region 14 corresponding to amino acids 302 to 338 of SEQ IDNO: 2, region 15 corresponding to amino acids 362 to 546 of SEQ ID NO:2, region 16 corresponding to amino acids 596 to 611 of SEQ ID NO: 2,region 17 corresponding to amino acids 661 to 805 of SEQ ID NO: 2,region 18 corresponding to amino acids 829 to 838 of SEQ ID NO: 2, andregion 19 corresponding to amino acids 1043 to 1055 of SEQ ID NO: 2.

In one embodiment the present disclosure relates to a detergentcomposition comprising an endoglucanase variant of the disclosure havingan alteration (e.g., a substitution, deletion or insertion) at one ormore positions in a region selected from the group of regions 1-9 (e.g.,of SEQ ID NO: 2 or another parent endoglucanase), wherein in an aqueoussolution comprising a detergent component said region (e.g., of SEQ IDNO: 2 or another parent endoglucanase) is relatively more accessible tosaid detergent component than one or more or all of its adjacentregions.

In one embodiment the present disclosure relates to a detergentcomposition comprising an endoglucanase variant of the disclosure havingan alteration (e.g., a substitution, deletion or insertion) at one ormore positions in a region selected from the group of regions 1-9 (e.g.,of SEQ ID NO: 2 or another parent endoglucanase), wherein in an aqueoussolution comprising a detergent component said region (e.g., of SEQ IDNO: 2 or another parent endoglucanase) is relatively more exposed tosaid detergent component than one or more or all of its adjacentregions.

In one embodiment the present disclosure relates to a detergentcomposition comprising an endoglucanase variant of the disclosure havingan alteration (e.g., a substitution, deletion or insertion) at one ormore positions in a region selected from the group of regions 1-9 (e.g.,of SEQ ID NO: 2 or another parent endoglucanase), wherein in an aqueoussolution comprising a detergent component said region (e.g., of SEQ IDNO: 2 or another parent endoglucanase) is relatively more accessible tosaid detergent component than one or more or all of its adjacentregions.

In one embodiment the present disclosure relates to a detergentcomposition comprising an endoglucanase variant of the disclosure havingan alteration (e.g., a substitution, deletion or insertion) at one ormore positions in a region selected from the group of regions 1-9 (e.g.,of SEQ ID NO: 2 or another parent endoglucanase), wherein in an aqueoussolution comprising a detergent component said region (e.g., of SEQ IDNO: 2 or another parent endoglucanase) is relatively moreconformationally dynamic than one or more or all of its adjacentregions.

In one embodiment the present disclosure relates to a detergentcomposition comprising an endoglucanase variant of the disclosure havingan alteration (e.g., a substitution, deletion or insertion) at one ormore positions in a region selected from the group of regions 1-9 (e.g.,of SEQ ID NO: 2 or another parent endoglucanase), wherein in an aqueoussolution comprising a detergent component said region (e.g., of SEQ IDNO: 2 or another parent endoglucanase) is relatively more receptive todeuterium incorporation than one or more or all of its adjacent regions.

In one embodiment the present disclosure relates to a detergentcomposition comprising an endoglucanase variant of the disclosure,further comprising an alteration (e.g., a substitution, deletion orinsertion) at one or more positions in one or more regions selected fromthe group of: region 1 corresponding to amino acids 95 to 105 of SEQ IDNO: 2, region 2 corresponding to amino acids 115 to 138 of SEQ ID NO: 2,region 3 corresponding to amino acids 210 to 251 of SEQ ID NO: 2, region4 corresponding to amino acids 267 to 301 of SEQ ID NO: 2, region 5corresponding to amino acids 339 to 361 of SEQ ID NO: 2, region 6corresponding to amino acids 547 to 595 of SEQ ID NO: 2, region 7corresponding to amino acids 612 to 660 of SEQ ID NO: 2, region 8corresponding to amino acids 806 to 828 of SEQ ID NO: 2, and region 9corresponding to amino acids 839 to 1042 of SEQ ID NO: 2, wherein saidvariant has at least about 60% and less than about 100% sequenceidentity to SEQ ID NO: 2, preferably said endoglucanase variant hasactivity on xanthan gum pretreated with xanthan lyase, furtherpreferably said activity is a xanthan gum degrading activity.

In one embodiment the present disclosure relates to a detergentcomposition comprising an endoglucanase variant of the disclosure havingmultiple alterations (such as 2, 3, 4, 5, 6, 7, 8, 9 or 10) in oneregion (e.g., of SEQ ID NO: 2 or another parent endoglucanase) selectedfrom the group of: region 1 corresponding to amino acids 95 to 105 ofSEQ ID NO: 2, region 2 corresponding to amino acids 115 to 138 of SEQ IDNO: 2, region 3 corresponding to amino acids 210 to 251 of SEQ ID NO: 2,region 4 corresponding to amino acids 267 to 301 of SEQ ID NO: 2, region5 corresponding to amino acids 339 to 361 of SEQ ID NO: 2, region 6corresponding to amino acids 547 to 595 of SEQ ID NO: 2, region 7corresponding to amino acids 612 to 660 of SEQ ID NO: 2, region 8corresponding to amino acids 806 to 828 of SEQ ID NO: 2, and region 9corresponding to amino acids 839 to 1042 of SEQ ID NO: 2, wherein saidvariant has at least about 60% and less than about 100% sequenceidentity to SEQ ID NO: 2, preferably said endoglucanase variant hasactivity on xanthan gum pretreated with xanthan lyase, furtherpreferably said activity is a xanthan gum degrading activity.

In one embodiment the present disclosure relates to a detergentcomposition comprising an endoglucanase variant of the disclosure havingmultiple alterations (e.g., about 2, 3, 4, 5, 6, 7, 8, 9 or 10) inmultiple regions (e.g., about 2, 3, 4, 5, 6, 7, 8, 9 or 10) (e.g., ofSEQ ID NO: 2 or another parent endoglucanase) selected from the groupof: region 1 corresponding to amino acids 95 to 105 of SEQ ID NO: 2,region 2 corresponding to amino acids 115 to 138 of SEQ ID NO: 2, region3 corresponding to amino acids 210 to 251 of SEQ ID NO: 2, region 4corresponding to amino acids 267 to 301 of SEQ ID NO: 2, region 5corresponding to amino acids 339 to 361 of SEQ ID NO: 2, region 6corresponding to amino acids 547 to 595 of SEQ ID NO: 2, region 7corresponding to amino acids 612 to 660 of SEQ ID NO: 2, region 8corresponding to amino acids 806 to 828 of SEQ ID NO: 2, and region 9corresponding to amino acids 839 to 1042 of SEQ ID NO: 2, wherein saidvariant has at least about 60% and less than about 100% sequenceidentity to SEQ ID NO: 2, preferably said endoglucanase variant hasactivity on xanthan gum pretreated with xanthan lyase, furtherpreferably said activity is a xanthan gum degrading activity.

In a further embodiment, the present disclosure relates to a detergentcomposition comprising an endoglucanase variant of the disclosure havingmultiple alterations (e.g., about 2, 3, 4, 5, 6, 7, 8, 9 or 10) inmultiple regions (e.g., about 2, 3, 4, 5, 6, 7, 8, 9 or 10) (e.g., ofSEQ ID NO: 2 or another parent endoglucanase) selected from the groupof: region 1 corresponding to amino acids 95 to 105 of SEQ ID NO: 2,region 2 corresponding to amino acids 115 to 138 of SEQ ID NO: 2, region3 corresponding to amino acids 210 to 251 of SEQ ID NO: 2, region 4corresponding to amino acids 267 to 301 of SEQ ID NO: 2, region 5corresponding to amino acids 339 to 361 of SEQ ID NO: 2, region 6corresponding to amino acids 547 to 595 of SEQ ID NO: 2, region 7corresponding to amino acids 612 to 660 of SEQ ID NO: 2, region 8corresponding to amino acids 806 to 828 of SEQ ID NO: 2, and region 9corresponding to amino acids 839 to 1042 of SEQ ID NO: 2, and multiplealterations (e.g. about 2, 3, 4, 5, 6, 7, 8, 9, or 10) in multipleadjacent regions (e.g. about 2, 3, 4, 5, 6, 7, 8, 9, or 10) selectedfrom the group of: region 10 corresponding to amino acids 1 to 94 of SEQID NO: 2, region 11 corresponding to amino acids 106 to 114 of SEQ IDNO: 2, region 12 corresponding to amino acids 139 to 209 of SEQ ID NO:2, region 13 corresponding to amino acids 252 to 266 of SEQ ID NO: 2,region 14 corresponding to amino acids 302 to 338 of SEQ ID NO: 2,region 15 corresponding to amino acids 362 to 546 of SEQ ID NO: 2,region 16 corresponding to amino acids 596 to 611 of SEQ ID NO: 2,region 17 corresponding to amino acids 661 to 805 of SEQ ID NO: 2,region 18 corresponding to amino acids 829 to 838 of SEQ ID NO: 2, andregion 19 corresponding to amino acids 1043 to 1055 of SEQ ID NO: 2;wherein said variant has at least about 60% and less than about 100%sequence identity to SEQ ID NO: 2, preferably said endoglucanase varianthas activity on xanthan gum pretreated with xanthan lyase, furtherpreferably said activity is a xanthan gum degrading activity.

In one embodiment, the present disclosure relates to a detergentcomposition comprising endoglucanase variants, comprising an alteration(e.g., a substitution, deletion or insertion) at one or more (e.g.,several) positions of the mature parent polypeptide (e.g., SEQ ID NO:2), wherein each alteration is independently a substitution, insertionor deletion, wherein the variant has endoglucanase activity.

In an embodiment, the variant has sequence identity of at least about60%, e.g., at least about 65%, at least about 70%, at least about 75%,at least about 80%, at least about 85%, at least about 90%, at leastabout 91%, at least about 92%, at least about 93%, at least about 94%,at least about 95%, at least about 96%, at least about 97%, at leastabout 98%, or at least about 99%, but less than about 100%, to the aminoacid sequence of the parent endoglucanase. In one embodiment, thevariant has at least about 60%, e.g., at least about 65%, at least about70%, at least about 75%, at least about 80%, at least about 85%, atleast about 90%, at least about 91%, at least about 92%, at least about93%, at least about 94%, at least about 95%, such as at least about 96%,at least about 97%, at least about 98%, or at least about 99%, but lessthan about 100%, sequence identity to the mature polypeptide of SEQ IDNO: 2.

In one embodiment the present disclosure relates to a detergentcomposition comprising an endoglucanase variant of the disclosure,wherein said variant has at least 61%, at least about 62%, at leastabout 63%, at least about 64%, at least about 65%, at least about 66%,at least about 67%, at least about 68%, at least about 69%, at leastabout 70%, at least about 71%, at least about 72%, at least about 73%,at least about 74%, at least about 75%, at least about 76%, at leastabout 77%, at least about 78%, at least about 79%, at least about 80%,at least about 81%, at least about 82%, at least about 83%, at leastabout 84%, at least about 85%, at least about 86%, at least about 87%,at least about 88%, at least about 89%, at least about 90%, at leastabout 91%, at least about 92%, at least about 93%, at least about 94%,at least about 95%, at least about 96%, at least about 97%, at leastabout 98%, or at least about 99% sequence identity to SEQ ID NO: 2.

In another aspect, a variant comprises an alteration at one or more(e.g., several) positions corresponding to positions 4, 17, 18, 20, 51,53, 55, 56, 60, 63, 71, 79, 87, 92, 99, 120, 125, 126, 130, 137, 182,186, 189, 192, 213, 216, 221, 226, 228, 230, 231, 232, 233, 235, 240,243, 247, 249, 278, 279, 281, 283, 285, 289, 292, 294, 298, 302, 311,313, 333, 346, 353, 358, 386, 387, 388, 390, 403, 408, 410, 416, 441,448, 451, 471, 472, 476, 489, 507, 512, 515, 538, 555, 556, 557, 558,559, 560, 561, 562, 563, 564, 567, 568, 570, 575, 578, 579, 580, 581,583, 589, 590, 591, 592, 593, 595, 598, 599, 602, 603, 605, 607, 609,616, 627, 630, 631, 635, 636, 638, 639, 640, 641, 642, 643, 644, 651,676, 683, 688, 690, 694, 698, 699, 706, 711, 713, 1719, 720, 744, 749,754, 756, 760, 781, 786, 797, 810, 811, 812, 815, 823, 824, 825, 827,828, 833, 834, 835, 837, 843, 848, 868, 869, 870, 871, 872, 873, 874,880, 881, 883, 884, 885, 887, 888, 890, 892, 894, 898, 905, 906, 912,920, 921, 924, 926, 927, 928, 932, 933, 934, 935, 937, 938, 939, 940,941, 942, 943, 946, 948, 950, 952, 953, 954, 956, 957, 960, 966, 971,972, 980, 989, 991, 994, 995, 998, 999, 1006, 1009, 1010, 1011, 1029,1030, 1031, 1032, 1035, 1037, 1038, 1040, 1041, 1042, 1044, 1045, 1048,wherein numbering is according to SEQ ID NO: 2.

In another aspect, a variant comprises an alteration at two positionscorresponding to any of positions 4, 17, 18, 20, 51, 53, 55, 56, 60, 63,71, 79, 87, 92, 99, 120, 125, 126, 130, 137, 182, 186, 189, 192, 213,216, 221, 226, 228, 230, 231, 232, 233, 235, 240, 243, 247, 249, 278,279, 281, 283, 285, 289, 292, 294, 298, 302, 311, 313, 333, 346, 353,358, 386, 387, 388, 390, 403, 408, 410, 416, 441, 448, 451, 471, 472,476, 489, 507, 512, 515, 538, 555, 556, 557, 558, 559, 560, 561, 562,563, 564, 567, 568, 570, 575, 578, 579, 580, 581, 583, 589, 590, 591,592, 593, 595, 598, 599, 602, 603, 605, 607, 609, 616, 627, 630, 631,635, 636, 638, 639, 640, 641, 642, 643, 644, 651, 676, 683, 688, 690,694, 698, 699, 706, 711, 713, 1719, 720, 744, 749, 754, 756, 760, 781,786, 797, 810, 811, 812, 815, 823, 824, 825, 827, 828, 833, 834, 835,837, 843, 848, 868, 869, 870, 871, 872, 873, 874, 880, 881, 883, 884,885, 887, 888, 890, 892, 894, 898, 905, 906, 912, 920, 921, 924, 926,927, 928, 932, 933, 934, 935, 937, 938, 939, 940, 941, 942, 943, 946,948, 950, 952, 953, 954, 956, 957, 960, 966, 971, 972, 980, 989, 991,994, 995, 998, 999, 1006, 1009, 1010, 1011, 1029, 1030, 1031, 1032,1035, 1037, 1038, 1040, 1041, 1042, 1044, 1045, 1048, wherein numberingis according to SEQ ID NO: 2.

In another aspect, a variant comprises an alteration at three positionscorresponding to any of positions 4, 17, 18, 20, 51, 53, 55, 56, 60, 63,71, 79, 87, 92, 99, 120, 125, 126, 130, 137, 182, 186, 189, 192, 213,216, 221, 226, 228, 230, 231, 232, 233, 235, 240, 243, 247, 249, 278,279, 281, 283, 285, 289, 292, 294, 298, 302, 311, 313, 333, 346, 353,358, 386, 387, 388, 390, 403, 408, 410, 416, 441, 448, 451, 471, 472,476, 489, 507, 512, 515, 538, 555, 556, 557, 558, 559, 560, 561, 562,563, 564, 567, 568, 570, 575, 578, 579, 580, 581, 583, 589, 590, 591,592, 593, 595, 598, 599, 602, 603, 605, 607, 609, 616, 627, 630, 631,635, 636, 638, 639, 640, 641, 642, 643, 644, 651, 676, 683, 688, 690,694, 698, 699, 706, 711, 713, 1719, 720, 744, 749, 754, 756, 760, 781,786, 797, 810, 811, 812, 815, 823, 824, 825, 827, 828, 833, 834, 835,837, 843, 848, 868, 869, 870, 871, 872, 873, 874, 880, 881, 883, 884,885, 887, 888, 890, 892, 894, 898, 905, 906, 912, 920, 921, 924, 926,927, 928, 932, 933, 934, 935, 937, 938, 939, 940, 941, 942, 943, 946,948, 950, 952, 953, 954, 956, 957, 960, 966, 971, 972, 980, 989, 991,994, 995, 998, 999, 1006, 1009, 1010, 1011, 1029, 1030, 1031, 1032,1035, 1037, 1038, 1040, 1041, 1042, 1044, 1045, 1048, wherein numberingis according to SEQ ID NO: 2.

In another aspect, a variant comprises an alteration at each position(or at least four positions) corresponding to positions 4, 17, 18, 20,51, 53, 55, 56, 60, 63, 71, 79, 87, 92, 99, 120, 125, 126, 130, 137,182, 186, 189, 192, 213, 216, 221, 226, 228, 230, 231, 232, 233, 235,240, 243, 247, 249, 278, 279, 281, 283, 285, 289, 292, 294, 298, 302,311, 313, 333, 346, 353, 358, 386, 387, 388, 390, 403, 408, 410, 416,441, 448, 451, 471, 472, 476, 489, 507, 512, 515, 538, 555, 556, 557,558, 559, 560, 561, 562, 563, 564, 567, 568, 570, 575, 578, 579, 580,581, 583, 589, 590, 591, 592, 593, 595, 598, 599, 602, 603, 605, 607,609, 616, 627, 630, 631, 635, 636, 638, 639, 640, 641, 642, 643, 644,651, 676, 683, 688, 690, 694, 698, 699, 706, 711, 713, 1719, 720, 744,749, 754, 756, 760, 781, 786, 797, 810, 811, 812, 815, 823, 824, 825,827, 828, 833, 834, 835, 837, 843, 848, 868, 869, 870, 871, 872, 873,874, 880, 881, 883, 884, 885, 887, 888, 890, 892, 894, 898, 905, 906,912, 920, 921, 924, 926, 927, 928, 932, 933, 934, 935, 937, 938, 939,940, 941, 942, 943, 946, 948, 950, 952, 953, 954, 956, 957, 960, 966,971, 972, 980, 989, 991, 994, 995, 998, 999, 1006, 1009, 1010, 1011,1029, 1030, 1031, 1032, 1035, 1037, 1038, 1040, 1041, 1042, 1044, 1045,1048, wherein numbering is according to SEQ ID NO: 2.

In another aspect, the variant comprises or consist of an alteration ata position corresponding to position 4. In another aspect, the aminoacid at a position corresponding to position 4 is substituted with Ala,Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro,Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises orconsists of the substitution V4T of the mature polypeptide of SEQ ID NO:2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 17. In another aspect, the aminoacid at a position corresponding to position 17 is substituted with Ala,Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro,Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises orconsists of the substitution S17A of the mature polypeptide of SEQ IDNO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 18. In another aspect, the aminoacid at a position corresponding to position 18 is substituted with Ala,Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro,Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises orconsists of the substitution N18G of the mature polypeptide of SEQ IDNO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 20. In another aspect, the aminoacid at a position corresponding to position 20 is substituted with Ala,Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro,Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises orconsist of the substitution F20P, F20N, F20G, or F20Y, preferably F20P,of the mature polypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 51. In another aspect, the aminoacid at a position corresponding to position 51 is substituted with Ala,Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro,Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises orconsists of the substitution K51Q or K51H, preferably K51Q, of themature polypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 53. In another aspect, the aminoacid at a position corresponding to position 53 is substituted with Ala,Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro,Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises orconsists of the substitution E53Y, E53P, or E53G, preferably E53Y, ofthe mature polypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 55. In another aspect, the aminoacid at a position corresponding to position 55 is substituted with Ala,Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro,Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises orconsists of the substitution Y55M or Y55D, preferably Y55M, of themature polypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 56. In another aspect, the aminoacid at a position corresponding to position 56 is substituted with Ala,Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro,Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises orconsists of the substitution V56M of the mature polypeptide of SEQ IDNO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 60. In another aspect, the aminoacid at a position corresponding to position 60 is substituted with Ala,Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro,Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises orconsists of the substitution Y60F of the mature polypeptide of SEQ IDNO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 63. In another aspect, the aminoacid at a position corresponding to position 63 is substituted with Ala,Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro,Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises orconsists of the substitution S63F of the mature polypeptide of SEQ IDNO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 71. In another aspect, the aminoacid at a position corresponding to position 71 is substituted with Ala,Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro,Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises orconsists of the substitution A71E of the mature polypeptide of SEQ IDNO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 79. In another aspect, the aminoacid at a position corresponding to position 79 is substituted with Ala,Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro,Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises orconsists of the substitution S79W of the mature polypeptide of SEQ IDNO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 87. In another aspect, the aminoacid at a position corresponding to position 87 is substituted with Ala,Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro,Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises orconsists of the substitution T87R of the mature polypeptide of SEQ IDNO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 92. In another aspect, the aminoacid at a position corresponding to position 92 is substituted with Ala,Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro,Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises orconsists of the substitution T92S of the mature polypeptide of SEQ IDNO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 120. In another aspect, the aminoacid at a position corresponding to position 120 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution A120P of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 129. In another aspect, the aminoacid at a position corresponding to position 129 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution N129D of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 137. In another aspect, the aminoacid at a position corresponding to position 137 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution F137L of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 182. In another aspect, the aminoacid at a position corresponding to position 182 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution H182Y of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 186. In another aspect, the aminoacid at a position corresponding to position 186 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution A186P of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 189. In another aspect, the aminoacid at a position corresponding to position 189 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution N189K of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 192. In another aspect, the aminoacid at a position corresponding to position 192 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution K192N of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 216. In another aspect, the aminoacid at a position corresponding to position 216 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution N216D, N216Q, r N216R,preferably N216D, of the mature polypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 226. In another aspect, the aminoacid at a position corresponding to position 226 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution L226K of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 228. In another aspect, the aminoacid at a position corresponding to position 228 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution K228E of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 230. In another aspect, the aminoacid at a position corresponding to position 230 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution G230H of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 233. In another aspect, the aminoacid at a position corresponding to position 233 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution L233H of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 247. In another aspect, the aminoacid at a position corresponding to position 247 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution D247N of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 278. In another aspect, the aminoacid at a position corresponding to position 278 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution A278S of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 279. In another aspect, the aminoacid at a position corresponding to position 279 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution G279E of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 281. In another aspect, the aminoacid at a position corresponding to position 281 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution K281R of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 283. In another aspect, the aminoacid at a position corresponding to position 283 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution A283D of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 285. In another aspect, the aminoacid at a position corresponding to position 285 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution N285L, N285M, N285S, N285P,N285T, N285Y, N285H, N285K, N285D, N285W, N285R, or N285G, preferablyN285G, of the mature polypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 289. In another aspect, the aminoacid at a position corresponding to position 289 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution Q289E of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 292. In another aspect, the aminoacid at a position corresponding to position 292 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution T292F, T292L, T292I, T292V,T292S, T292P, T292Y, T292Q, T292N, T292K, T292D, T292A, or T292G,preferably T292A, of the mature polypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 294. In another aspect, the aminoacid at a position corresponding to position 294 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution A294V of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 297. In another aspect, the aminoacid at a position corresponding to position 297 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution F297L of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 298. In another aspect, the aminoacid at a position corresponding to position 298 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution Q298E of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 302. In another aspect, the aminoacid at a position corresponding to position 302 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution I302D, I302H, I302V, or I302M,preferably I302D, of the mature polypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 311. In another aspect, the aminoacid at a position corresponding to position 311 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution H311N of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 313. In another aspect, the aminoacid at a position corresponding to position 313 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution S313D of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 333. In another aspect, the aminoacid at a position corresponding to position 333 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution W333L of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 346. In another aspect, the aminoacid at a position corresponding to position 346 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution A346H or A246D of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 353. In another aspect, the aminoacid at a position corresponding to position 353 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution T353D of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 386. In another aspect, the aminoacid at a position corresponding to position 386 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution A386P of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 387. In another aspect, the aminoacid at a position corresponding to position 387 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution I387T of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 388. In another aspect, the aminoacid at a position corresponding to position 388 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution K388R of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 390. In another aspect, the aminoacid at a position corresponding to position 390 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution K390Q of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 403. In another aspect, the aminoacid at a position corresponding to position 403 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution I403Y of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 408 In another aspect, the aminoacid at a position corresponding to position 408 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution E408D, E408N, E408S, E408P,E408A, E408G, or E408G, preferably E408D, of the mature polypeptide ofSEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 410. In another aspect, the aminoacid at a position corresponding to position 410 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution P410G of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 416. In another aspect, the aminoacid at a position corresponding to position 416 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution Q416S or Q416D, preferablyQ416S, of the mature polypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 441. In another aspect, the aminoacid at a position corresponding to position 441 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution N441G of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 448. In another aspect, the aminoacid at a position corresponding to position 448 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution A448E, A448W, or A448S,preferably A448E, of the mature polypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 451. In another aspect, the aminoacid at a position corresponding to position 451 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution K451S or K451Q, preferablyK451S, of the mature polypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 471. In another aspect, the aminoacid at a position corresponding to position 471 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution G471S of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 472. In another aspect, the aminoacid at a position corresponding to position 472 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution S472Y of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 476. In another aspect, the aminoacid at a position corresponding to position 476 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution D476R of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 489. In another aspect, the aminoacid at a position corresponding to position 489 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution Q489P of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 507. In another aspect, the aminoacid at a position corresponding to position 507 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution K507R of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 512. In another aspect, the aminoacid at a position corresponding to position 512 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution K512P of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 515. In another aspect, the aminoacid at a position corresponding to position 515 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution S515V of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 538. In another aspect, the aminoacid at a position corresponding to position 538 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution S538C of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 555. In another aspect, the aminoacid at a position corresponding to position 555 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution L555Q of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 557. In another aspect, the aminoacid at a position corresponding to position 557 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution G557R of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 558. In another aspect, the aminoacid at a position corresponding to position 558 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the alteration N558D, N558NP, N558F, N558I,N558E, or N558M of the mature polypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 559. In another aspect, the aminoacid at a position corresponding to position 559 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution A559S, A559N, A559F, A559M,A559P, A559Y, A559H, A559Q, A559D, or A559G, preferably A559N, of themature polypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 560. In another aspect, the aminoacid at a position corresponding to position 560 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution S560P and S560G, preferablyS560P, of the mature polypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 561. In another aspect, the aminoacid at a position corresponding to position 561 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution T561P of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 564. In another aspect, the aminoacid at a position corresponding to position 564 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution A564I, A564Y, A564Q, A564E, orA564K, preferably A564I, of the mature polypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 567. In another aspect, the aminoacid at a position corresponding to position 567 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution V567F or V567P of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 568. In another aspect, the aminoacid at a position corresponding to position 568 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution K568R of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 570. In another aspect, the aminoacid at a position corresponding to position 570 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution P570K, P570Q, P570R, P570T,P570S, P570A, P570H, P570G, and P570N, preferably P570K or P570R, of themature polypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 575. In another aspect, the aminoacid at a position corresponding to position 575 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution I575V of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 579. In another aspect, the aminoacid at a position corresponding to position 579 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution Y579W or Y579F, preferablyY579W, of the mature polypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 581. In another aspect, the aminoacid at a position corresponding to position 581 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution T581M of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 592. In another aspect, the aminoacid at a position corresponding to position 592 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution G592D of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 593. In another aspect, the aminoacid at a position corresponding to position 593 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution S593N and S593E, preferablyS593N, of the mature polypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 595. In another aspect, the aminoacid at a position corresponding to position 595 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution S595L of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 598. In another aspect, the aminoacid at a position corresponding to position 598 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution S598Q of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 599. In another aspect, the aminoacid at a position corresponding to position 599 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution A599S of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or of an alteration at aposition corresponding to position 602. In another aspect, the aminoacid at a position corresponding to position 602 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or of the substitution I602T or I602D, preferably I602T, ofthe mature polypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 603. In another aspect, the aminoacid at a position corresponding to position 603 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution V603P of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 605. In another aspect, the aminoacid at a position corresponding to position 605 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution S605T of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 607. In another aspect, the aminoacid at a position corresponding to position 607 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution S607C of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 609. In another aspect, the aminoacid at a position corresponding to position 609 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution G609E of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 616. In another aspect, the aminoacid at a position corresponding to position 616 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution S616D or S616G, preferablyS616D, of the mature polypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 627. In another aspect, the aminoacid at a position corresponding to position 627 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution K627L, K627M, K627V, K627S,K627T, K627Q, or K627R, preferably K627R, of the mature polypeptide ofSEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 630. In another aspect, the aminoacid at a position corresponding to position 630 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution I630F, I630V, or I630Y of themature polypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 631. In another aspect, the aminoacid at a position corresponding to position 631 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution K631A or K631R, preferablyK631R, of the mature polypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 633. In another aspect, the aminoacid at a position corresponding to position 633 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution T633V of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 635. In another aspect, the aminoacid at a position corresponding to position 635 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution D635P, D635N, D635K, D635E,D635W, D635L, D635M, D635T, D635A, or D635G, preferably D635A, of themature polypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 636. In another aspect, the aminoacid at a position corresponding to position 636 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution S636M, S636A, S636H, S636Q,S636N, S636R, S636L, S636H, or S636K, preferably S636N, of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 638. In another aspect, the aminoacid at a position corresponding to position 638 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution F638N, F638I, F638V, F638T,F638L, F638Y, F638M or F638H, preferably F638N, of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 639. In another aspect, the aminoacid at a position corresponding to position 639 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution T639S, T639I, T639M, T639V,T639A, T639D, T639E, T639Y, T639W, T639P, or T639G, preferably T639G orT639I, of the mature polypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 640. In another aspect, the aminoacid at a position corresponding to position 640 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution T640S of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 642. In another aspect, the aminoacid at a position corresponding to position 642 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution S642T or S642N, preferablyS642N, of the mature polypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 643. In another aspect, the aminoacid at a position corresponding to position 643 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution N643D or N643H, preferablyN643D, of the mature polypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 651. In another aspect, the aminoacid at a position corresponding to position 651 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution A651P or A651S, preferablyA651P, of the mature polypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 676. In another aspect, the aminoacid at a position corresponding to position 676 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution D676H of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 683. In another aspect, the aminoacid at a position corresponding to position 683 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution Q683E of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 688. In another aspect, the aminoacid at a position corresponding to position 688 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution A688G of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 690. In another aspect, the aminoacid at a position corresponding to position 690 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution Y690F of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 694. In another aspect, the aminoacid at a position corresponding to position 694 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution T694A of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 697. In another aspect, the aminoacid at a position corresponding to position 697 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution T697G of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 698. In another aspect, the aminoacid at a position corresponding to position 698 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution R698W of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 699. In another aspect, the aminoacid at a position corresponding to position 699 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution T699A of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 706. In another aspect, the aminoacid at a position corresponding to position 706 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution T706Q of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 711. In another aspect, the aminoacid at a position corresponding to position 711 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution T711S, T711V, or T711Y,preferably T711V, of the mature polypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 713. In another aspect, the aminoacid at a position corresponding to position 713 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution K713R of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 719. In another aspect, the aminoacid at a position corresponding to position 719 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution W719R of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 720. In another aspect, the aminoacid at a position corresponding to position 720 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution K720H of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 744. In another aspect, the aminoacid at a position corresponding to position 744 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution K744H or K744Q, preferablyK744H, of the mature polypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 749. In another aspect, the aminoacid at a position corresponding to position 749 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution A749T of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 754. In another aspect, the aminoacid at a position corresponding to position 754 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution K754R of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 756. In another aspect, the aminoacid at a position corresponding to position 756 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution V756Y or V756H, preferablyV756Y, of the mature polypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 760. In another aspect, the aminoacid at a position corresponding to position 760 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution S760G of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 781. In another aspect, the aminoacid at a position corresponding to position 781 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution T781M of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 786. In another aspect, the aminoacid at a position corresponding to position 786 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution N786K of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 797. In another aspect, the aminoacid at a position corresponding to position 797 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution T797S of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 810. In another aspect, the aminoacid at a position corresponding to position 810 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution S810Q of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 824. In another aspect, the aminoacid at a position corresponding to position 824 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution A824D of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 825. In another aspect, the aminoacid at a position corresponding to position 825 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution T825G of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 828. In another aspect, the aminoacid at a position corresponding to position 828 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution N828D of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 833. In another aspect, the aminoacid at a position corresponding to position 833 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution N833D of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 834. In another aspect, the aminoacid at a position corresponding to position 834 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution Q834E of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 835. In another aspect, the aminoacid at a position corresponding to position 835 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution S835A or S835D, preferablyS835A, of the mature polypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 837. In another aspect, the aminoacid at a position corresponding to position 837 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution V837I of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 848. In another aspect, the aminoacid at a position corresponding to position 848 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution N848D of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 868. In another aspect, the aminoacid at a position corresponding to position 868 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution A868E of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 869. In another aspect, the aminoacid at a position corresponding to position 869 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution A869V of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 870. In another aspect, the aminoacid at a position corresponding to position 870 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution D870V of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 872. In another aspect, the aminoacid at a position corresponding to position 872 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution T872G, T872H, T872W, or T872Q,preferably T872G, of the mature polypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 880. In another aspect, the aminoacid at a position corresponding to position 880 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution R880K of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 881. In another aspect, the aminoacid at a position corresponding to position 881 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution V881Q or V881T, preferablyV881Q, of the mature polypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 883. In another aspect, the aminoacid at a position corresponding to position 883 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution T883R, T883V, T883C, or T883K,preferably T883R, of the mature polypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 884. In another aspect, the aminoacid at a position corresponding to position 884 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution Y884H of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 885. In another aspect, the aminoacid at a position corresponding to position 885 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution A885Q, A885N, or A885F,preferably A885F, of the mature polypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 887. In another aspect, the aminoacid at a position corresponding to position 887 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution T887S or T887K, preferablyT887K, of the mature polypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 888. In another aspect, the aminoacid at a position corresponding to position 888 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution L888M of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 890. In another aspect, the aminoacid at a position corresponding to position 890 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution V890R of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 892. In another aspect, the aminoacid at a position corresponding to position 892 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution T892V or T892P, preferablyT892P, of the mature polypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 898. In another aspec, the aminoacid at a position corresponding to position 898 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution R898Q of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 905. In another aspect, the aminoacid at a position corresponding to position 905 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution N905D of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 906. In another aspect, the aminoacid at a position corresponding to position 906 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution F906A of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 912. In another aspect, the aminoacid at a position corresponding to position 912 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution Q912V of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 920. In another aspect, the aminoacid at a position corresponding to position 920 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution N920D or N920P of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 921. In another aspect, the aminoacid at a position corresponding to position 921 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution K921R or K921E of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 924. In another aspect, the aminoacid at a position corresponding to position 924 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution A924D of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 926. In another aspect, the aminoacid at a position corresponding to position 926 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution V926F or V926P, preferablyV926P, of the mature polypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 927. In another aspect, the aminoacid at a position corresponding to position 927 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution K927R of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 928. In another aspect, the aminoacid at a position corresponding to position 928 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution S928D of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 932. In another aspect, the aminoacid at a position corresponding to position 932 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution T932A of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 933. In another aspect, the aminoacid at a position corresponding to position 933 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution N933S or N933V, preferablyN933S, of the mature polypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 934. In another aspect, the aminoacid at a position corresponding to position 934 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution Y934G, Y034R, or Y934Q,preferably Y934G, of the mature polypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 937. In another aspect, the aminoacid at a position corresponding to position 937 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution A937E of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 938. In another aspect, the aminoacid at a position corresponding to position 938 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution V938I of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 939. In another aspect, the aminoacid at a position corresponding to position 939 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution K939V of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 941. In another aspect, the aminoacid at a position corresponding to position 941 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution N941S of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 942. In another aspect, the aminoacid at a position corresponding to position 942 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or of the substitution A942P of the mature polypeptide of SEQID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 946. In another aspect, the aminoacid at a position corresponding to position 946 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution G946R of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 948. In another aspect, the aminoacid at a position corresponding to position 948 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution K948R of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 956. In another aspect, the aminoacid at a position corresponding to position 956 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution Q956Y or A956S, preferablyQ956Y, of the mature polypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 957. In another aspect, the aminoacid at a position corresponding to position 957 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution A957L or A957P, preferablyA957L, of the mature polypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 966. In another aspect, the aminoacid at a position corresponding to position 966 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution N966C of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 972. In another aspect, the aminoacid at a position corresponding to position 972 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution T972K of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 980. In another aspect, the aminoacid at a position corresponding to position 980 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution M980I of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 994. In another aspect, the aminoacid at a position corresponding to position 994 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution G994N or G994D of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 999. In another aspect, the aminoacid at a position corresponding to position 999 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution T999R of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 1011. In another aspect, the aminoacid at a position corresponding to position 1011 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution L1011A of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 1031. In another aspect, the aminoacid at a position corresponding to position 1031 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution K1031I of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 1037. In another aspect, the aminoacid at a position corresponding to position 1037 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution A1037E of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 1038. In another aspect, the aminoacid at a position corresponding to position 1038 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution S1038G of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 1041. In another aspect, the aminoacid at a position corresponding to position 1041 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution G1041R of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 1042. In another aspect, the aminoacid at a position corresponding to position 1042 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution Y1042N of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position 1048. In another aspect, the aminoacid at a position corresponding to position 1048 is substituted withAla, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe,Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variantcomprises or consists of the substitution F1048W of the maturepolypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of alterations atpositions corresponding to positions 559+579. In another aspect, theamino acids at positions corresponding to positions 559+579 areindependently substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly,His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. Inanother aspect, the variant comprises or consists of the substitutionsA559N+Y579W or A559N+Y579F of the mature polypeptide of SEQ ID NO: 2.

In another aspect, the variant comprises or consists of an alteration ata position corresponding to position selected from the group ofalterations in positions: 4, 17, 18, 20, 51, 53, 55, 56, 60, 63, 71, 79,87, 92, 99, 120, 125, 126, 130, 137, 182, 186, 189, 192, 213, 216, 221,226, 228, 230, 231, 232, 233, 235, 240, 243, 247, 249, 278, 279, 281,283, 285, 289, 292, 294, 298, 302, 311, 313, 333, 346, 353, 358, 386,387, 388, 390, 403, 408, 410, 416, 441, 448, 451, 471, 472, 476, 489,507, 512, 515, 538, 555, 556, 557, 558, 559, 560, 561, 562, 563, 564,567, 568, 570, 575, 578, 579, 580, 581, 583, 589, 590, 591, 592, 593,595, 598, 599, 602, 603, 605, 607, 609, 616, 627, 630, 631, 635, 636,638, 639, 640, 641, 642, 643, 644, 651, 676, 683, 688, 690, 694, 698,699, 706, 711, 713, 1719, 720, 744, 749, 754, 756, 760, 781, 786, 797,810, 811, 812, 815, 823, 824, 825, 827, 828, 833, 834, 835, 837, 843,848, 868, 869, 870, 871, 872, 873, 874, 880, 881, 883, 884, 885, 887,888, 890, 892, 894, 898, 905, 906, 912, 920, 921, 924, 926, 927, 928,932, 933, 934, 935, 937, 938, 939, 940, 941, 942, 943, 946, 948, 950,952, 953, 954, 956, 957, 960, 966, 971, 972, 980, 989, 991, 994, 995,998, 999, 1006, 1009, 1010, 1011, 1029, 1030, 1031, 1032, 1035, 1037,1038, 1040, 1041, 1042, 1044, 1045, 1048, of SEQ ID NO: 2.

In another aspect, the amino acid at a position corresponding to any ofpositions as described above is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val. In another aspect, the variant comprises or consists of thesubstitution selected from the group of: N285G, W333L, T353D, N558NP,N558F, T633V, D635L, D635M, D635T, F638Y, T639D, G994N, and K281T,G563E, I575M, I575A, K921D, N558K+A559K+S560F+T561P+G562W, N558K, A559K,S560F, T561P, G562W and I125V, A126R, K130R, K213R, A221R, K228E, K228I,G230F, G230L, G230A, G230H, G230N, G230W, G230T, F231Y, F231N, V232R,V232G, H235D, N240Q, G243K, G243R, A249N, A278S, K281F, K281V, K281Y,K281H, K281Q, K281N, K281W, N285L, N285M, N285S, N285P, N285T, N285Y,N285H, N285K, N285D, N285W, N285R, T292F, T292L, T292I, T292V, T292S,T292P, T292Y, T292Q, T292N, T292K, T292D, T292G, F297L, A346H, G556S,N558D, N558M, N558Q, N558I, N558Y, N558H, A559N, A559F, A559M, A559P,A559Y, A559H, A559Q, A559D, A559R, A559G, A559I, A559S, S560P, S560K,S560G, S560D, T561P, T561E, T561Q, T561S, T561D, A564I, A564Y, A564H,A564Q, A564K, A564E, E565M, V567F, K568R, L569F, L569Y, L569D, L569E,P570F, P570L, P570I, P570M, P570V, P570S, P570T, P570A, P570Y, P570H,P570Q, P570N, P570K, P570E, P570W, P570R, P570G, I575D, I575E, I576F,I576M, I576P, D578R, Y579F, Y579W, V580L, D583M, Q589G, P590S, P590T,P590E, E591L, G592D, S593P, S593H, S593Q, S593N, S593K, S593D, S593E,S593R, S616D, K627L, K627M, K627V, K627S, K627T, K627Q, K627R, I630F,I630V, I630Y, D635A, D635P, D635N, D635K, D635E, D635G, D635W, S636L,S636M, S636A, S636H, S636Q, S636N, S636K, S636R, F638I, F638V, F638T,F638L, F638H, T639V, T639S, T639L, T639I, T639M, T639A, T639E, T639W,T639G, Y641E, S642T, S642N, N643D, N643H, N643T, T644F, A651P, S810R,A811S, V812F, V812I, V812M, V812W, V812R, N815V, N815Y, N815E, N815W,N815R, S823Q, A824T, T825N, T825W, T825A, T825D, V827I, V827M, V827S,T843V, D870F, D870L, D870I, D870M, D870V, D870S, D870T, D870Y, D870H,D870Q, D870N, D870K, D870E, D870W, D870R, D870G, P871F, P871L, P871I,P871M, P871V, P871S, P871T, P871A, P871Y, P871H, P871Q, T872S, T872F,T872A, T872Y, T872H, T872Q, T872N, T872K, T872D, T872E, T872W, T872R,T872G, D873K, D873E, T874V, T874S, T874P, T874A, T874H, T874Q, T874N,T874K, V881Q, T883K, Y884H, A885F, A885Q, A885N, T887L, T887I, T887S,T887H, T887R, K894E, N920D, K921R, K921E, T932A, N933V, N933S, Y934G,Y934M, Y934S, Y934A, Y934Q, Y934N, Y934E, Y934W, Y934R, T935W, A937F,A937V, A937S, A937T, A937Q, A937D, A937E, V938I, K939I, K939V, D940E,N941S, N941H, N941D, A942P, A942E, D943Y, D943H, R950V, R950H, R950N,F952S, F952W, N953Y, G954L, Y960F, A964N, A964C, N966P, N966C, G971A,Q974K, Q974C, Q989I, Q991L, Q991I, Q991M, Q991V, Q991T, Q991K, Q991C,S995I, S995V, S995Q, S995R, S995C, G998V, G998A, S1006T, S1006A, S1006K,S1006R, Y1010W, L1011M, L1011S, L1011A, L1011Q, L1011N, L1011D, L1011E,R1029N, F1030M, K1031I, K1031S, K1031T, K1031H, V1032G, K1035A, A1037E,A1037W, S1038L, S1038I, L1040N, L1040E, G1041F, L1044F, L1044S, L1044N,L1044W, P1045Q, P1045W, and A559N+Y579F, A559N, Y579F, A564E+Y579F,A564E, Y579F, A559N+Y579W, A559N, Y579W, G562P+Y579W, G562P, Y579W,A564D+Y579W, A564D, Y579W, A559N+Y579W+K99R, A559N, Y579W, K99R,A559N+Y579W+K281R, A559N, Y579W, K281R, K281R+A559N+Y579W, K281R, A559N,Y579W, A559N+Y579W+S616D, A559N, Y579W, S616D, A559N+Y579W+S636N, A559N,Y579W, S636N, A559N+Y579W+A651P, A559N, Y579W, A651P, A559N+Y579W+K948E,A559N, Y579W, K948E, A559N+Y579W+K1009E, A559N, Y579W, K1009E,A559N+Y579W+K627R, A559N, Y579W, K627R, Y579W+K921R, Y579W, K921R,A559N+Y579W+K921R, A559N, Y579W, K921R, K99R+Y579W, K99R, Y579W,Y579W+A651P, Y579W, A651P, Y579W+K948E, Y579W, K948E, Y579W+K1009E,Y579W, K1009E, A559N+Y579W+Y934G, A559N, Y579W, Y934G,A559N+Y579W+K921R+Y934G, A559N, Y579W, K921R, Y934G, A559N+Y579W+K627M,A559N, Y579W, K627M, A559N+Y579W+K627R+S616D, A559N, Y579W, K627R,S616D, A559N+Y579F+K627R, A559N, Y579F, K627R, A559N+Y579W+K921R+A651P,A559N, Y579W, K921R, A651P, A559N+Y579W+K921R+K627R, A559N, Y579W,K921R, K627R, A559N+Y579W+K921R+S636K, A559N, Y579W, K921R, S636K,A559N+Y579W+K921R+S616D, A559N, Y579W, K921R, S616D,A559N+Y579W+K921R+S636N, A559N, Y579W, K921R, S636N,A559N+Y579W+K921R+K627R+S636N, A559N, Y579W, K921R, K627R, S636N,A559N+Y579W+S636N+A651P, A559N, Y579W, S636N, A651P,A559N+Y579W+S616D+A651P, A559N, Y579W, S616D, A651P,A559N+Y579W+S616D+S636K, A559N, Y579W, S616D, S636K,A559N+Y579W+S616D+K921R+Y934G, A559N, Y579W, S616D, K921R, Y934G,A559N+Y579W+A651P+K627M, A559N, Y579W, A651P, K627M,A559N+Y579W+A651P+S636K, A559N, Y579W, A651P, S636K,A559N+Y579W+A651P+K627R+S636N, A559N, Y579W, A651P, K627R, S636N,A559N+Y579W+A651P+S616D, A559N, Y579W, A651P, S616D,A559N+Y579W+A651P+K921R+Y934G, A559N, Y579W, A651P, K921R, Y934G,S636N+Y934G, S636N, Y934G, S636N+K921R, S636N, K921R, S636N+K627R,S636N, K627R, S636N+Y579W, S636N, Y579W, F638I+Y934G, F638I, Y934G,F638I+K921R, F638I, K921R, F638I+K627R, F638I, K627R, F638I+Y579W,F638I, Y579W, K627R+K51Q, K627R, K51Q, K627R+K451S, K627R, K451S,K627R+A559N, K627R, A559N, K627R+Y579W, K627R, Y579W, Y579W+Y934G,Y579W, Y934G, A651P+F638I, A651P, F638I, P570Q+A651P, P570Q, A651P,P570Q+K921R, P570Q, K921R, P570Q+K627R, P570Q, K627R, P570Q+A559N,P570Q, A559N, P570Q+Y579W, P570Q, Y579W, P570Q+F638I, P570Q, F638I,P570K+Y579W, P570K, Y579W, P570K+F638I, P570K, F638I, P570T+A651P,P570T, A651P, P570T+S636N, P570T, S636N, P570T+Y934G, P570T, Y934G,P570T+F638I, P570T, F638I, P570T+K921R, P570T, K921R, P570T+K627R,P570T, K627R, P570T+A559N, P570T, A559N, P570T+A885F, P570T, A885F,A885F+Y934G, A885F, Y934G, A885F+K627R, A885F, K627R, A559N+Y579W+S636L,A559N, Y579W, S636L, A559N+Y579W+F638I, A559N, Y579W, F638I,A559N+Y579W+D870M, D870M, A559N+Y579W+S560P, S560P, A559N+Y579W+A564I,A564I, A559N+Y579W+P570N, P570N, A559N+Y579W+P570K, P570K,A559N+Y579W+P570R, P570R, A559N+Y579W+P570A, P570A, A559N+Y579W+P570T,P570T, A559N+Y579W+P570S, P570S, A559N+Y579W+P570Q, P570Q,A559N+Y579W+P570H, P570H, and N558E, A559P, A559N, A559H, T561P, A564E,P570A, P570Q, P570R, P570S, P570K, P570T, P570N, Y579W, Y579F, T581M,S616D, K627R, K627M, K627Q, S636N, S636Q, S636R, S636K, S636M, S636H,F638I, F638L, N643D, A651P, A651S, A885F, A885Q, K921R, Y934R, Y934G,N966C, L1011A, K1031I, and A559N+P570A+Y579W, A559N+P570H+Y579W,A559N+P570K+Y579W, A559N+P570N+Y579W, A559N+P570Q+Y579W,A559N+P570R+Y579W, A559N+P570S+Y579W, A559N+P570T+Y579W,A559N+S560P+Y579W, A559N+Y579W+A651P, A559N+Y579W+A651P+Y934G,A559N+Y579W+F638I, A559N+Y579W+K921R, A559N+Y579W+S616D+K921R,A559N+Y579W+S636N, A559N+Y579F, A559N+Y579W, A559N+Y579W+K921R,A559N+Y579W+S616D, F638I+Y934G, K627R+S636N, K627R+Y934G, P570K+Y579W,Q416D+A559N+Y579W+S636N, Q416D, S128X+A559N+Y579W+K627R, S128X,S128X+A559N+Y579W+S636N, Y579W+S636N, V4T, S17A, N18G, F20P, F20N, F20G,F20Y, K51Q, K51H, E53Y, E53P, E53G, Y55M, Y55D, V56M, Y60F, S63F, A71E,579W, T87R, T92S, A120P, N129D, F137L, H182Y, A186P, N189K, K192N,N216D, N216Q, N216R, L226K, G230H, L233H, D247N, G279E, K281R, A283D,N285D, N285G, Q289E, T292A, T292F, T292Y, A294V, Q298E, I302D, I302H,I302V, I302M, H311N, S313D, A346D, A386P, I387T, K388R, K390Q, I403Y,E408D, E408N, E408S, E408P, E408A, E408G, P410G, Q416S, Q416D, N441G,A448E, A448W, A448S, K451S, K451Q, G471S, S472Y, D476R, Q489P, K507R,K512P, S515V, S538C, L555Q, G557R, N558E, A559N, A559P, A559H, A559D,S560P, S560G, T561P, A564E, A564I, V567P, K568R, P570R, P570Q, P570K,P570A, P570T, P570G, P570S, P570H, P570N, I575V, Y579W, Y579F, T581M,S593N, S593E, S595L, S598Q, A599S, I602T, I602D, V603P, S605T, S607C,G609E, S616G, S616D, K627R, K627M, K627Q, K631R, K631A, D635A, D635E,D635M, D635N, D635L, D635W, S636N, S636K, S636L, S636Q, S636R, S636M,S636H, F638N, F638I, F638L, F638V, F638H, F638M, T639G, T639I, T639M,T639Y, T639W, T639P, T639E, T640S, S642N, S642T, N643D, N643H, A651P,A651S, D676H, Q683E, A688G, Y690F, T694A, T697G, R698W, T699A, T706Q,T711S, T711V, T711Y, K713R, W719R, K720H, K744H, K744Q, A749T, K754R,V756Y, V756H, S760G, T781M, N786K, T797S, S810Q, A824D, T825G, N828D,N833D, Q834E, S835A, S835D, V837I, N848D, A868E, A869V, D870V, T872G,T872H, T872W, T872Q, R880K, V881Q, V881T, T883R, T883V, T883C, T883K,Y884H, A885N, A885Q, A885F, T887K, T887S, L888M, V890R, T892P, T892V,R898Q, N905D, F906A, Q912V, N920P, K921R, A924D, V926F, V926P, K927R,S928D, T932A, N933S, N933V, Y934G, Y934R, Y934Q, A937E, V938I, K939V,N941S, A942P, G946R, K948R, Q956Y, Q956S, A957L, A957P, N966C, T972K,M980I, G994D, T999R, L1011A, K1031I, A1037E, S1038G, G1041R, Y1042N, andF1048W, wherein numbering is according to SEQ ID NO: 2.

The variants may further comprise one or more additional alterations atone or more (e.g., several) other positions.

The amino acid changes may be of a minor nature, that is conservativeamino acid substitutions or insertions that do not significantly affectthe folding and/or activity of the protein; small deletions, typicallyof about 1-30 amino acids; small amino- or carboxyl-terminal extensions,such as an amino-terminal methionine residue; a small linker peptide ofup to about 20-25 residues; or a small extension that facilitatespurification by changing net charge or another function, such as apoly-histidine tract, an antigenic epitope or a binding domain.

Examples of conservative substitutions are within the groups of basicamino acids (arginine, lysine and histidine), acidic amino acids(glutamic acid and aspartic acid), polar amino acids (glutamine andasparagine), hydrophobic amino acids (leucine, isoleucine and valine),aromatic amino acids (phenylalanine, tryptophan and tyrosine), and smallamino acids (glycine, alanine, serine, threonine and methionine). Aminoacid substitutions that do not generally alter specific activity areknown in the art and are described, for example, by H. Neurath and R. L.Hill, 1979, In, The Proteins, Academic Press, New York. Commonsubstitutions are Ala/Ser, Val/Ile, Asp/Glu, Thr/Ser, Ala/Gly, Ala/Thr,Ser/Asn, Ala/Val, Ser/Gly, Tyr/Phe, Ala/Pro, Lys/Arg, Asp/Asn, Leu/Ile,Leu/Val, Ala/Glu, and Asp/Gly.

Alternatively, the amino acid changes are of such a nature that thephysico-chemical properties of the polypeptides are altered. Forexample, amino acid changes may improve the thermal stability of thepolypeptide, alter the substrate specificity, change the pH optimum, andthe like.

Essential amino acids in a polypeptide can be identified according toprocedures known in the art, such as site-directed mutagenesis oralanine-scanning mutagenesis (Cunningham and Wells, 1989, Science 244:1081-1085). In the latter technique, single alanine mutations areintroduced at every residue in the molecule, and the resultant mutantmolecules are tested for xanthan lyase activity to identify amino acidresidues that are critical to the activity of the molecule. See also,Hilton et al., 1996, J. Biol. Chem. 271: 4699-4708. The active site ofthe enzyme or other biological interaction can also be determined byphysical analysis of structure, as determined by such techniques asnuclear magnetic resonance, crystallography, electron diffraction, orphotoaffinity labeling, in conjunction with mutation of putative contactsite amino acids. See, for example, de Vos et al., 1992, Science 255:306-312; Smith et al., 1992, J. Mol. Biol. 224: 899-904; Wlodaver etal., 1992, FEBS Lett. 309: 59-64. The identity of essential amino acidscan also be inferred from an alignment with a related polypeptide.

In one embodiment, the present disclosure relates to a detergentcomposition comprising an endoglucanase variant of the disclosure,having the total number of alterations compared to SEQ ID NO: 2 between1 and 20, e.g., between 1 and 10 or between 1 and 5, such as about 1, 2,3, 4, 5, 6, 7, 8, 9 or 10 alterations.

In one embodiment, the present disclosure relates to a detergentcomposition comprising an endoglucanase variant of the disclosure,wherein an activity on xanthan gum pretreated with xanthan lyase is axanthan degrading activity, preferably said xanthan degrading activityis endoglucanase EC 3.2.1.4 activity; further preferably said xanthanlyase has EC 4.2.2.12 activity.

In an embodiment, the variant has an improved stability in a detergentcomposition compared to a parent enzyme (e.g., SEQ ID NO: 2). In oneembodiment, the improved stability is measured as an improved half-life.In one embodiment, the improved stability is measured as half-lifeimprovement factor.

In one embodiment, the present disclosure relates to a detergentcomposition comprising an endoglucanase variant of the disclosure,wherein said variant has an improved stability in a detergentcomposition compared to a parent endoglucanase (e.g., with SEQ ID NO:2); preferably said detergent composition comprises a chelator; furtherpreferably said chelator is EDTA or citrate.

In one embodiment, the present disclosure relates to a detergentcomposition comprising an endoglucanase variant of the disclosure,wherein said variant has a half-life improvement factor (HIF)≥1.0;preferably said variant has a half-life improvement factor (HIF)>1.0. Apreferred way of calculating said half-life improvement factor (HIF) isdescribed in example 4 herein. Accordingly, residual activity (RA) canbe calculated using the following formula:

${R\;{A(\%)}} = {\frac{{Abs}({Stress})}{{Abs}({Ref})} \times 100\%}$wherein Abs(Stress) is the absorbance at about 405 nm of the sample inthe stress microtiter plate (MTP) (e.g. incubated at about 25° C.over-night or any other combination of temperature and time) aftersubtracting relevant background absorbance contributions, Abs(Ref) isthe absorbance at 405 nm of the sample in the reference MTP (e.g.incubated at about 4° C. over-night) after subtracting relevantbackground absorbance contributions, wherein half-lives for thedegradation of each variant and parent endoglucanase (e.g. at about 25°C.) are calculated using the following formula:

${T\;{1/2}} = {- \frac{{\ln(2)} \times T}{\ln( \frac{{Abs}({Stress})}{{Abs}({Ref})} )}}$wherein T is the incubation time for both the stress and reference MTP,wherein half-life-improvement factors (HIFs) are calculated using thefollowing formula:

${HIF} = \frac{{T\;{1/2}},{variant}}{{T\;{1/2}},{wt}}$e.g. wherein T½ wt (or wild type) is the T½ of the mature parentendoglucanase with SEQ ID NO: 2.

In one embodiment the present disclosure relates to a detergentcomposition comprising an endoglucanase variant of the disclosure,wherein a half-life improvement factor (HIF) is determined afterincubation of said endoglucanase variant in a detergent composition atabout 25° C. for a time period from about 17 to about 20 hours.

Parent

The parent endoglucanase may be (a) a polypeptide having at least 60%sequence identity to the mature polypeptide of SEQ ID NO: 2; (b) apolypeptide encoded by a polynucleotide that hybridizes under lowstringency conditions with (i) the mature polypeptide coding sequence ofSEQ ID NO: 1, or (ii) the full-length complement of (i); or (c) apolypeptide encoded by a polynucleotide having at least 60% sequenceidentity to the mature polypeptide coding sequence of SEQ ID NO: 1.

In an aspect, the parent has a sequence identity to the maturepolypeptide of SEQ ID NO: 2 of at least about 60%, e.g., at least about65%, at least about 70%, at least about 75%, at least about 80%, atleast about 85%, at least about 90%, at least about 91%, at least about92%, at least about 93%, at least about 94%, at least about 95%, atleast about 96%, at least about 97%, at least about 98%, at least about99%, or about 100%, which have xanthan lyase activity. In one aspect,the amino acid sequence of the parent differs by up to 10 amino acids,e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, from the mature polypeptide ofSEQ ID NO: 2.

In another aspect, the parent comprises or consists of the amino acidsequence of SEQ ID NO: 2. In another aspect, the parent comprises orconsists of the mature polypeptide of SEQ ID NO: 2. In another aspect,the parent is a fragment of the mature polypeptide of SEQ ID NO: 2containing at least about 85%, about 86%, about 87%, about 88%, about89%, about 90%, about 91%, about 92%, about 93%, about 94% or about 95%of the number of amino acids of SEQ ID NO: 2. In another embodiment, theparent is an allelic variant of the mature polypeptide of SEQ ID NO: 2.

In another aspect, the parent is encoded by a polynucleotide thathybridizes under very low stringency conditions, low stringencyconditions, medium stringency conditions, medium-high stringencyconditions, high stringency conditions, or very high stringencyconditions with (i) the mature polypeptide coding sequence of SEQ ID NO:1, or (ii) the full-length complement of (i) (Sambrook et al., 1989,Molecular Cloning, A Laboratory Manual, 2d edition, Cold Spring Harbor,N.Y.).

The polynucleotide of SEQ ID NO: 1 or a subsequence thereof, as well asthe polypeptide of SEQ ID NO: 2 or a fragment thereof, may be used todesign nucleic acid probes to identify and clone DNA encoding a parentfrom strains of different genera or species according to methods wellknown in the art. In particular, such probes can be used forhybridization with the genomic DNA or cDNA of a cell of interest,following standard Southern blotting procedures, in order to identifyand isolate the corresponding gene therein. Such probes can beconsiderably shorter than the entire sequence, but should be at leastabout 15, e.g., at least about 25, at least about 35, or at least about70 nucleotides in length. Preferably, the nucleic acid probe is at leastabout 100 nucleotides in length, e.g., at least about 200 nucleotides,at least about 300 nucleotides, at least about 400 nucleotides, at leastabout 500 nucleotides, at least about 600 nucleotides, at least about700 nucleotides, at least about 800 nucleotides, or at least about 900nucleotides in length. Both DNA and RNA probes can be used. The probesare typically labeled for detecting the corresponding gene (for example,with ³²P, ³H, ³⁵S, biotin, or avidin). Such probes are encompassed bythe present disclosure.

A genomic DNA or cDNA library prepared from such other strains may bescreened for DNA that hybridizes with the probes described above andencodes a parent. Genomic or other DNA from such other strains may beseparated by agarose or polyacrylamide gel electrophoresis, or otherseparation techniques. DNA from the libraries or the separated DNA maybe transferred to and immobilized on nitrocellulose or of her suitablecarrier material. In order to identify a clone or DNA that hybridizeswith SEQ ID NO: 1 or a subsequence thereof, the carrier material is usedin a Southern blot.

For purposes of the present disclosure, hybridization indicates that thepolynucleotide hybridizes to a labeled nucleic acid probe correspondingto (i) SEQ ID NO: 1; (ii) the mature polypeptide coding sequence of SEQID NO: 1; (iii) the full-length complement thereof, or (iv) asubsequence thereof, under very low to very high stringency conditions.Molecules to which the nucleic acid probe hybridizes under theseconditions can be detected using, for example, X-ray film or any otherdetection mechanism or apparatus known in the art.

In one aspect, the nucleic acid probe is the mature polypeptide codingsequence of SEQ ID NO: 1. In another aspect, the nucleic acid probe is apolynucleotide that encodes the polypeptide of SEQ ID NO: 2; the maturepolypeptide thereof; or a fragment thereof. In another aspect, thenucleic acid probe is SEQ ID NO: 1.

In another embodiment, the parent is encoded by a polynucleotide havinga sequence identity to the mature polypeptide coding sequence of SEQ IDNO: 1 of at least about 60%, e.g., at least about 65%, at least about70%, at least about 75%, at least about 80%, at least about 85%, atleast about 90%, at least about 91%, at least about 92%, at least about93%, at least about 94%, at least about 95%, at least about 96%, atleast about 97%, at least about 98%, at least about 99%, or about 100%.

The polypeptide may be a hybrid polypeptide in which a region of onepolypeptide is fused at the N-terminus or the C-terminus of a region ofanother polypeptide.

The parent may be a fusion polypeptide or cleavable fusion polypeptidein which another polypeptide is fused at the N-terminus or theC-terminus of the polypeptide of the present disclosure. A fusionpolypeptide is produced by fusing a polynucleotide encoding anotherpolypeptide to a polynucleotide of the present disclosure. Techniquesfor producing fusion polypeptides are known in the art, and includeligating the coding sequences encoding the polypeptides so that they arein frame and that expression of the fusion polypeptide is under controlof the same promoter(s) and terminator. Fusion polypeptides may also beconstructed using intein technology in which fusion polypeptides arecreated post-translationally (Cooper et al., 1993, EMBO J. 12:2575-2583; Dawson et al., 1994, Science 266: 776-779).

A fusion polypeptide can further comprise a cleavage site between thetwo polypeptides. Upon secretion of the fusion protein, the site iscleaved releasing the two polypeptides. Examples of cleavage sitesinclude, but are not limited to, the sites disclosed in Martin et al.,2003, J. Ind. Microbiol. Biotechnol. 3: 568-576; Svetina et al., 2000,J. Biotechnol. 76: 245-251; Rasmussen-Wilson et al., 1997, Appl.Environ. Microbiol. 63: 3488-3493; Ward et al., 1995, Biotechnology 13:498-503; and Contreras et al., 1991, Biotechnology 9: 378-381; Eaton etal., 1986, Biochemistry 25: 505-512; Collins-Racie et al., 1995,Biotechnology 13: 982-987; Carter et al., 1989, Proteins: Structure,Function, and Genetics 6: 240-248; and Stevens, 2003, Drug DiscoveryWorld 4: 35-48.

The parent may be obtained from microorganisms of any genus. Forpurposes of the present disclosure, the term “obtained from” as usedherein in connection with a given source shall mean that the parentencoded by a polynucleotide is produced by the source or by a strain inwhich the polynucleotide from the source has been inserted. In oneaspect, the parent is secreted extracellularly.

The parent may be a bacterial enzyme. For example, the parent may be aGram-positive bacterial polypeptide such as a Bacillus, Clostridium,Enterococcus, Geobacillus, Lactobacillus, Lactococcus, Oceanobacillus,Staphylococcus, Streptococcus, or Streptomyces enzyme, or aGram-negative bacterial polypeptide such as a Campylobacter, E. coli,Flavobacterium, Fusobacterium, Helicobacter, Ilyobacter, Neisseria,Pseudomonas, Salmonella, or Ureaplasma enzyme.

In one aspect, the parent is a Bacillus alkalophilus, Bacillusamyloliquefaciens, Bacillus brevis, Bacillus circulans, Bacillusclausii, Bacillus coagulans, Bacillus firmus, Bacillus lautus, Bacilluslentus, Bacillus licheniformis, Bacillus megaterium, Bacillus pumilus,Bacillus stearothermophilus, Bacillus subtilis, or Bacillusthuringiensis enzyme.

In another aspect, the parent is a Streptococcus equisimilis,Streptococcus pyogenes, Streptococcus uberis, or Streptococcus equisubsp. Zooepidemicus enzyme.

In another aspect, the parent is a Streptomyces achromogenes,Streptomyces avermitilis, Streptomyces coelicolor, Streptomyces griseus,or Streptomyces lividans enzyme.

The parent may be a fungal enzyme. For example, the parent may be ayeast enzyme such as a Candida, Kluyveromyces, Pichia, Saccharomyces,Schizosaccharomyces, or Yarrowia enzyme; or a filamentous fungal enzymesuch as an Acremonium, Agaricus, Alternaria, Aspergillus, Aureobasidium,Botryosphaeria, Ceriporiopsis, Chaetomidium, Chrysosporium, Claviceps,Cochliobolus, Coprinopsis, Coptotermes, Corynascus, Cryphonectria,Cryptococcus, Diplodia, Exidia, Filibasidium, Fusarium, Gibberella,Holomastigotoides, Humicola, Irpex, Lentinula, Leptospaeria,Magnaporthe, Melanocarpus, Meripilus, Mucor, Myceliophthora,Neocallimastix, Neurospora, Paecilomyces, Penicillium, Phanerochaete,Piromyces, Poitrasia, Pseudoplectania, Pseudotrichonympha, Rhizomucor,Schizophyllum, Scytalidium, Talaromyces, Thermoascus, Thielavia,Tolypocladium, Trichoderma, Trichophaea, Verticillium, Volvariella, orXylaria enzyme.

In another aspect, the parent is a Saccharomyces carlsbergensis,Saccharomyces cerevisiae, Saccharomyces diastaticus, Saccharomycesdouglasii, Saccharomyces kluyveri, Saccharomyces norbensis, orSaccharomyces oviformis enzyme.

In another aspect, the parent is an Acremonium cellulolyticus,Aspergillus aculeatus, Aspergillus awamori, Aspergillus foetidus,Aspergillus fumigatus, Aspergillus japonicus, Aspergillus nidulans,Aspergillus niger, Aspergillus oryzae, Chrysosporium inops,Chrysosporium keratinophilum, Chrysosporium lucknowense, Chrysosporiummerdarium, Chrysosporium pannicola, Chrysosporium queenslandicum,Chrysosporium tropicum, Chrysosporium zonatum, Fusarium bactridioides,Fusarium cerealis, Fusarium crookwellense, Fusarium culmorum, Fusariumgraminearum, Fusarium graminum, Fusarium heterosporum, Fusarium negundi,Fusarium oxysporum, Fusarium reticulatum, Fusarium roseum, Fusariumsambucinum, Fusarium sarcochroum, Fusarium sporotrichioides, Fusariumsulphureum, Fusarium torulosum, Fusarium trichothecioides, Fusariumvenenatum, Humicola grisea, Humicola insolens, Humicola lanuginosa,Irpex lacteus, Mucor miehei, Myceliophthora thermophila, Neurosporacrassa, Penicillium funiculosum, Penicillium purpurogenum, Phanerochaetechrysosporium, Thielavia achromatica, Thielavia albomyces, Thielaviaalbopilosa, Thielavia australeinsis, Thielavia fimeti, Thielaviamicrospora, Thielavia ovispora, Thielavia peruviana, Thielavia setosa,Thielavia spededonium, Thielavia sub thermophila, Thielavia terrestris,Trichoderma harzianum, Trichoderma koningii, Trichodermalongibrachiatum, Trichoderma reesei, or Trichoderma viride enzyme.

In another aspect, the parent is a Paenibacillus sp. xanthan lyase,e.g., the xanthan lyase of SEQ ID NO: 2.

It will be understood that for the aforementioned species, thedisclosure encompasses both the perfect and imperfect states, and othertaxonomic equivalents, e.g., anamorphs, regardless of the species nameby which they are known. Those skilled in the art will readily recognizethe identity of appropriate equivalents.

Strains of these species are readily accessible to the public in anumber of culture collections, such as the American Type CultureCollection (ATCC), Deutsche Sammlung von Mikroorganismen andZellkulturen GmbH (DSMZ), Centraalbureau Voor Schimmelcultures (CBS),and Agricultural Research Service Patent Culture Collection, NorthernRegional Research Center (NRRL).

The parent may be identified and obtained from other sources includingmicroorganisms isolated from nature (e.g., soil, composts, water, etc.)or DNA samples obtained directly from natural materials (e.g., soil,composts, water, etc.) using the above-mentioned probes. Techniques forisolating microorganisms and DNA directly from natural habitats are wellknown in the art. A polynucleotide encoding a parent may then beobtained by similarly screening a genomic DNA or cDNA library of anothermicroorganism or mixed DNA sample. Once a polynucleotide encoding aparent has been detected with the probe(s), the polynucleotide can beisolated or cloned by utilizing techniques that are known to those ofordinary skill in the art (see, e.g., Sambrook et al., 1989, supra).

Preparation of Variants

Methods for obtaining a variant having endoglucanase activity, maycomprise: (a) introducing into a parent endoglucanase an alteration atone or more (e.g., several) positions corresponding to positions 4, 17,18, 20, 51, 53, 55, 56, 60, 63, 71, 79, 87, 92, 99, 120, 125, 126, 130,137, 182, 186, 189, 192, 213, 216, 221, 226, 228, 230, 231, 232, 233,235, 240, 243, 247, 249, 278, 279, 281, 283, 285, 289, 292, 294, 298,302, 311, 313, 333, 346, 353, 358, 386, 387, 388, 390, 403, 408, 410,416, 441, 448, 451, 471, 472, 476, 489, 507, 512, 515, 538, 555, 556,557, 558, 559, 560, 561, 562, 563, 564, 567, 568, 570, 575, 578, 579,580, 581, 583, 589, 590, 591, 592, 593, 595, 598, 599, 602, 603, 605,607, 609, 616, 627, 630, 631, 635, 636, 638, 639, 640, 641, 642, 643,644, 651, 676, 683, 688, 690, 694, 698, 699, 706, 711, 713, 1719, 720,744, 749, 754, 756, 760, 781, 786, 797, 810, 811, 812, 815, 823, 824,825, 827, 828, 833, 834, 835, 837, 843, 848, 868, 869, 870, 871, 872,873, 874, 880, 881, 883, 884, 885, 887, 888, 890, 892, 894, 898, 905,906, 912, 920, 921, 924, 926, 927, 928, 932, 933, 934, 935, 937, 938,939, 940, 941, 942, 943, 946, 948, 950, 952, 953, 954, 956, 957, 960,966, 971, 972, 980, 989, 991, 994, 995, 998, 999, 1006, 1009, 1010,1011, 1029, 1030, 1031, 1032, 1035, 1037, 1038, 1040, 1041, 1042, 1044,1045, 1048, wherein numbering is according to SEQ ID NO: 2. wherein thevariant has endoglucanase activity; and (b) recovering the variant.

In an alternative embodiment, the present disclosure also relates tomethods for obtaining a variant having endoglucanase activity,comprising: (a) introducing into a parent endoglucanase an alteration atone or more (e.g., several) positions corresponding to positions 285,333, 353, 558, 633, 635, 638, 639, 994, 281, 563, 575, 921,558+559+560+561+562, 558, 559, 560, 561, 562 125, 126, 130, 213, 221,228, 230, 231, 232, 235, 240, 243, 249, 278, 292, 297, 346, 556, 564,565, 567, 568, 569, 570, 576, 578, 579, 580, 583, 589, 590, 591, 592,593, 616, 627, 630, 636, 641, 642, 643, 644, 651, 810, 811, 812, 815,823, 824, 825, 827, 843, 870, 871, 872, 873, 874, 881, 883, 884, 885,887, 894, 920, 932, 933, 934, 935, 937, 938, 939, 940, 941, 942, 943,950, 952, 953, 954, 960, 964, 966, 971, 974, 989, 991, 995, 998, 1006,1010, 1011, 1029, 1030, 1031, 1032, 1035, 1037, 1038, 1040, 1041, 1044,1045, 559+579, 564+579, 562+579, 559+579+99, 99, 559+579+281,281+559+579, 559+579+616, 559+579+636, 559+579+651, 559+579+948, 948,559+579+1009, 1009, 559+579+627, 579+921, 559+579+921, 99+579, 579+651,579+948, 579+1009, 559+579+934, 934, 559+579+921+934, 559+579+627,559+579+627+616, 559+579+627, 559+579+921+651, 559+579+921+627,559+579+921+636, 559+579+921+616, 559+579+921+636, 559+579+921+627+636,559+579+636+651, 559+579+616+651, 559+579+616+636, 559+579+616+921+934,559+579+651+627, 559+579+651+636, 559+579+651+627+636, 559+579+651+616,559+579+651+921+934, 636+934, 636+921, 636+627, 636+579, 638+934,638+921, 638+627, 638+579, 627+51, 51, 627+451, 451, 627+559, 627+579,579+934, 651+638, 570+651, 570+921, 570+627, 570+559, 570+579, 570+638,570+579, 570+638, 570+651, 570+636, 570+934, 570+638, 570+921, 570+627,570+559, 570+885, 885+934, 885+627, 559+579+636, 559+579+638,559+579+870, 559+579+560, 559+579+564, 559+579+570, 559+579+570,559+579+570, 559+579+570, 559+579+570, 559+579+570, 559+579+570,559+579+570, 559+570+579, 559+570+579, 559+570+579, 559+570+579,559+570+579, 559+570+579, 559+570+579, 559+570+579, 559+560+579,559+579+651, 559+579+651+934, 559+579+638, 559+579+921, 559+579+616+921,559+579+636, 559+579, 559+579, 559+579+921, 559+579+616, 638+934,627+636, 627+934, 570+579, 416+559+579+636, 128+559+579+627,128+559+579+636, and 579+636 of SEQ ID NO: 2, wherein the variant hasendoglucanase activity; and (b) recovering the variant.

In one embodiment, the variant has a half-life improvement factor (HIF)of at least about 10, such as at least about 20, such as at least about50, or such as at least about 100. In a particular embodiment, thevariant comprises or consists of the alterations selected from the groupof:

-   -   F20P+I302D+S313D+E408D+D476R+Y579W+S636K+T697G+V756Y+V881Q+T887K+F906A+A937E;    -   F20P+I302D+S313D+E408D+D476R+Y579W+S636K+T697G+V756Y+V881Q+T887K+F906A+A937E;    -   F20P+I302D+S313D+E408D+D476R+Y579W+S636K+T697G+V756Y+V881Q+T887K+F906A+A937E;    -   F20P+I302D+S313D+E408D+D476R+Y579W+S636K+T697G+V756Y+T887K+F906A+A937E;    -   S17A+F20P+N216D+A283D+H311N+E408D+Y579W+I602T+A651P+A688G+T883R+F906A+Y934G+Q956Y;    -   F20P+I302D+S313D+E408D+D476R+Y579W+S636K+A651P+T697G+W719R+V756Y+N848D+V881T+T887K+F906A+A937E;    -   F20P+I302D+S313D+E408D+D476R+Y579W+S636K+A651P+T697G+W719R+V756Y+N848D+V881T+T887K+F906A+A937E;    -   F20P+I302D+S313D+E408D+D476R+Y579W+S636K+A651P+T697G+W719R+V756Y+N848D+V881T+T887K+F906A+A937E;    -   F20P+I302D+S313D+E408D+D476R+Y579W+S636K+A651P+T697G+W719R+V756Y+N848D+V881T+T887K+F906A+A937E;    -   F20P+S313D+E408D+Y579W+S616G+S636K+T697G+W719R+T883V+N905D+A        937E    -   F20P+S313D+E408D+Y579W+S636K+A688G+T697G+N905D+A937E;    -   F20P+I302D+S313D+E408D+D476R+Y579W+S636K+T697G+W719R+V756Y+V881Q+T887K+F906A+A937E;    -   F20P+N216Q+I302D+S313D+E408D+V567P+Y579W+S636K+A651P+T697G+W719R+N848D+T883R+N905D+A937E+Q956Y+F1048W;    -   F20P+K51Q+I302D+S313D+E408D+D476R+Q489P+A559N+Y579W+I602T+S636K+A651P+T697G+W719R+V756Y+N848D+T883R+T887K+F906A+A937E;    -   F20P+K51Q+I302D+S313D+E408D+D476R+Q489P+A559N+Y579W+I602T+S636K+A651P+T697G+W719R+V756Y+N848D+T883R+T887K+F906A+A937E;    -   F20P+I302D+S313D+E408D+D476R+Q489P+A559N+Y579W+I602T+S636K+A651P+T697G+W719R+V756Y+N848D+T883R+T887K+F906A+A937E;    -   F20P+I302D+S313D+E408D+D476R+Q489P+A559N+Y579W+I602T+S636K+A651P+T697G+W719R+V756Y+N848D+T883R+T887K+F906A+A937E;    -   F20P+I302D+S313D+A346D+E408D+D476R+Q489P+Y579W+S636N+T697G+W719R+V756Y+A824D+N848D+V881Q+T887K+F906A+S928D+A937E+F1048W;    -   F20P+I302D+S313D+A346D+E408D+D476R+Q489P+Y579W+S636N+T697G+W719R+V756Y+A824D+N848D+V881Q+T887K+F906A+S928D+A937E+F1048W;    -   N216D+S313D+E408D+D476R+Y579W+I602T+V603P+F638N+A651P+A688G+T697G+W719R+V756H+T825G+N833D+A869V+R880K+T887K+K921R+S928D+Y934G+N941S+K948R;    -   F20P+I302D+S313D+E408D+D476R+Q489P+Y579W+S636N+T697G+W719R+V756Y+A824D+V881Q+T887K+F906A+A937E+T999R+A1037E;    -   F20P+I302D+S313D+E408D+D476R+Q489P+Y579W+S636N+T697G+W719R+V756Y+A824D+V881Q+T887K+F906A+A937E+T999R+A1037E;    -   F20P+I302D+S313D+A346D+E408D+D476R+Y579W+S636N+T697G+W719R+V756Y+A824D+N848D+V881Q+T887K+F906A+A937E+T999R+F1048W;    -   F20P+I302D+S313D+A346D+E408D+D476R+Y579W+S636N+T697G+W719R+V756Y+A824D+N848D+V881Q+T887K+F906A+A937E+T999R+F1048W;    -   N216Q+S313D+E408D+D476R+Y579W+I602T+F638N+A651P+T697G+W719R+R880K+T887K+K921R+Y934G;    -   N216D+S313D+E408D+D476R+A564E+Y579W+I602T+F638N+A651P+Y690F+T697G+W719R+V756H+N833D+A869V+R880K+V881T+T887K+K921R+S928D+Y934G+T999R;    -   F20P+I302D+S313D+E408D+D476R+Y579W+S636K+T697G+W719R+V756Y+N848D+A869V+V881Q+T887K+N905D+F906A+Q912V+A937E+T999R+F1048W;    -   F20P+I302D+S313D+E408D+D476R+Q489P+Y579W+S636N+T697G+W719R+V756Y+A824D+N848D+V881Q+T887K+F906A+S928D+A937E;    -   F20P+I302D+S313D+E408D+D476R+Q489P+Y579W+S636N+T697G+W719R+V756Y+A824D+N848D+V881Q+T887K+F906A+S928D+A937E;    -   F20P+I302D+S313D+E408D+Q416S+D476R+Q489P+Y579W+I602T+S636K+A651P+T697G+W719R+V756Y+A824D+N833D+N848D+T883R+T887K+F906A+A937E;    -   F20P+I302D+S313D+E408D+Q416S+D476R+Q489P+Y579W+I602T+S636K+A651P+T697G+W719R+V756Y+A824D+N833D+N848D+T883R+T887K+F906A+A937E;    -   F20P+A186P+I302D+S313D+E408D+D476R+Q489P+Y579W+A599S+I602T+S636K+A651P+T697G+W719R+V756Y+N848D+T883R+T887K+F906A+A937E;    -   F20P+A186P+I302D+S313D+E408D+D476R+Q489P+Y579W+A599S+I602T+S636K+A651P+T697G+W719R+V756Y+N848D+T883R+T887K+F906A+A937E;    -   N216D+S313D+E408D+D476R+Y579W+I602T+V603P+F638N+A651P+A688G+T697G+W719R+V756H+R880K+T887K+K921R+S928D+Y934G+K948R;    -   F20P+K51Q+I302D+S313D+E408D+D476R+Q489P+Y579W+I602T+S636K+A651P+T697G+W719R+V756Y+N848D+T883R+T887K+F906A+A937E;    -   F20P+K51Q+I302D+S313D+E408D+D476R+Q489P+Y579W+I602T+S636K+A651P+T697G+W719R+V756Y+N848D+T883R+T887K+F906A+A937E;    -   F20P+I302D+S313D+A346D+E408D+D476R+Q489P+Y579W+S636N+T697G+W719R+V756Y+A824D+N848D+V881Q+T887K+F906A+A937E+T999R;    -   F20P+I302D+S313D+A346D+E408D+D476R+Q489P+Y579W+S636N+T697G+W719R+V756Y+A824D+N848D+V881Q+T887K+F906A+A937E+T999R;    -   F20P+I302D+S313D+E408D+D476R+Q489P+Y579W+I602T+S636N+T697G+W719R+V756Y+A824D+N848D+V881Q+T887K+N905D+F906A+A937E+T999R+A1037E+F1048W;    -   F20P+I302D+S313D+E408D+D476R+Q489P+Y579W+I602T+S636N+T697G+W719R+V756Y+A824D+N848D+V881Q+T887K+N905D+F906A+A937E+T999R+A1037E+F1048W;    -   F20P+K51Q+I302D+S313D+E408D+D476R+Q489P+Y579W+I602T+S636K+A651P+T697G+W719R+V756Y+A824D+N848D+T883R+T887K+F906A+S928D+A937E+A1037E;    -   F20P+K51Q+I302D+S313D+E408D+D476R+Q489P+Y579W+I602T+S636K+A651P+T697G+W719R+V756Y+A824D+N848D+T883R+T887K+F906A+S928D+A937E+A1        037E;    -   N216D+S313D+A346D+E408D+D476R+Q489P+A559P+Y579W+I602T+F638N+A651P+A688G+T6973+W719R+V756H+R880K+T887K+K921R+S928D+Y934G;    -   F20P+I302D+S313D+A346D+E408D+D476R+Q489P+Y579W+I602T+T697G+W719R+V756Y+N848D+V881Q+T887K+F906A+A937E;    -   F20P+I302D+S313D+A346D+E408D+D476R+Q489P+Y579W+I602T+T697G+W719R+V756Y+N848D+V881Q+T887K+F906A+A937E;    -   N216D+S313D+E408D+D476R+Q489P+A559P+Y579W+I602T+F638N+A651P+A        688G+T697G+W719R+V756H+Q834E+R880K+T887K+T892P+K921R+S928D+Y934G;    -   F20P+I302D+S313D+E408D+D476R+Q489P+A559N+Y579W+I602T+S636K+A651P+T697G+W719R+V756Y+N848D+T883R+T887K+F906A+S928D+A937E;    -   F20P+I302D+S313D+E408D+D476R+Q489P+A559N+Y579W+I602T+S636K+A651P+T697G+W719R+V756Y+N848D+T883R+T887K+F906A+S928D+A937E;    -   F20P+I302D+S313D+E408D+D476R+Q489P+Y579W+I602T+S636K+A651P+T697G+W719R+V756Y+N848D+T883R+T887K+F906A+A937E;    -   F20P+I302D+S313D+E408D+D476R+Q489P+Y579W+I602T+S636K+A651P+T697G+W719R+V756Y+N848D+T883R+T887K+F906A+A937E;    -   F20P+I302D+S313D+E408D+Q416S+D476R+Q489P+A559N+Y579W+I602T+S636K+A651P+T697G+W719R+V756Y+N848D+T883R+T887K+F906A+A937E;    -   F20P+I302D+S313D+E408D+Q416S+D476R+Q489P+A559N+Y579W+I602T+S636K+A651P+T697G+W719R+V756Y+N848D+T883R+T887K+F906A+A937E;    -   F20P+I302D+S313D+E408D+D476R+Y579W+I602T+S636N+T697G+W719R+V756Y+A824D+N848D+V881Q+T887K+F906A+S928D+A937E+T999R+F1048W;        and    -   F20P+I302D+S313D+E408D+D476R+Y579W+I602T+S636N+T697G+W719R+V756Y+A824D+N848D+V881Q+T887K+F906A+S928D+A937E+T999R+F1048W.

The variants can be prepared using any mutagenesis procedure known inthe art, such as site-directed mutagenesis, synthetic gene construction,semi-synthetic gene construction, random mutagenesis, shuffling, etc.

Site-directed mutagenesis is a technique in which one or more (e.g.,several) mutations are introduced at one or more defined sites in apolynucleotide encoding the parent.

Site-directed mutagenesis can be accomplished in vitro by PCR involvingthe use of oligonucleotide primers containing the desired mutation.Site-directed mutagenesis can also be performed in vitro by cassettemutagenesis involving the cleavage by a restriction enzyme at a site inthe plasmid comprising a polynucleotide encoding the parent andsubsequent ligation of an oligonucleotide containing the mutation in thepolynucleotide. Usually the restriction enzyme that digests the plasmidand the oligonucleotide is the same, permitting sticky ends of theplasmid and the insert to ligate to one another. See, e.g., Scherer andDavis, 1979, Proc. Natl. Acad. Sci. USA 76: 4949-4955; and Barton etal., 1990, Nucleic Acids Res. 18: 7349-4966.

Site-directed mutagenesis can also be accomplished in vivo by methodsknown in the art. See, e.g., U.S. Patent Application Publication No.2004/0171154; Storici et al., 2001, Nature Biotechnol. 19: 773-776; Krenet al., 1998, Nat. Med. 4: 285-290; and Calissano and Macino, 1996,Fungal Genet. Newslett. 43: 15-16.

Any site-directed mutagenesis procedure can be used in the presentdisclosure. There are many commercial kits available that can be used toprepare variants.

Synthetic gene construction entails in vitro synthesis of a designedpolynucleotide molecule to encode a polypeptide of interest. Genesynthesis can be performed utilizing a number of techniques, such as themultiplex microchip-based technology described by Tian et al. (2004,Nature 432: 1050-1054) and similar technologies wherein oligonucleotidesare synthesized and assembled upon photo-programmable microfluidicchips.

Single or multiple amino acid substitutions, deletions, and/orinsertions can be made and tested using known methods of mutagenesis,recombination, and/or shuffling, followed by a relevant screeningprocedure, such as those disclosed by Reidhaar-Olson and Sauer, 1988,Science 241: 53-57; Bowie and Sauer, 1989, Proc. Natl. Acad. Sci. USA86: 2152-2156; WO 95/17413; or WO 95/22625. Other methods that can beused include error-prone PCR, phage display (e.g., Lowman et al., 1991,Biochemistry 30: 10832-10837; U.S. Pat. No. 5,223,409; WO 92/06204) andregion-directed mutagenesis (Derbyshire et al., 1986, Gene 46: 145; Neret al., 1988, DNA 7: 127).

Mutagenesis/shuffling methods can be combined with high-throughput,automated screening methods to detect activity of cloned, mutagenizedpolypeptides expressed by host cells (Ness et al., 1999, NatureBiotechnology 17: 893-896). Mutagenized DNA molecules that encode activepolypeptides can be recovered from the host cells and rapidly sequencedusing standard methods in the art. These methods allow the rapiddetermination of the importance of individual amino acid residues in apolypeptide.

Semi-synthetic gene construction is accomplished by combining aspects ofsynthetic gene construction, and/or site-directed mutagenesis, and/orrandom mutagenesis, and/or shuffling. Semi-synthetic construction istypified by a process utilizing polynucleotide fragments that aresynthesized, in combination with PCR techniques. Defined regions ofgenes may thus be synthesized de novo, while other regions may beamplified using site-specific mutagenic primers, while yet other regionsmay be subjected to error-prone PCR or non-error prone PCRamplification. Polynucleotide subsequences may then be shuffled.

Embodiments

In one embodiment, the present disclosure relates to a detergentcomposition comprising at least one (e.g. about 1, 2, 3, 4, 5, 6, 7, 8,9 or 10) endoglucanase variant as described herein.

In one embodiment the present disclosure relates to a detergentcomposition comprising at least one (e.g. about 1, 2, 3, 4, 5, 6, 7, 8,9 or 10) endoglucanase variant as described herein, further comprisingone or more detergent components; preferably said detergent component isa chelator; further preferably said chelator is EDTA or citrate.

In one embodiment the present disclosure relates to a detergentcomposition comprising at least one (e.g. about 1, 2, 3, 4, 5, 6, 7, 8,9 or 10) endoglucanase variant as described herein, wherein saidcomposition is a detergent composition further comprising one or moredetergent components; preferably said detergent component comprises achelator; further preferably said chelator is EDTA or citrate.

In one embodiment, the present disclosure relates to a detergentcomposition comprising a composition comprising at least one (e.g. about1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) endoglucanase variant as describedherein, and further comprising one or more additional enzymes selectedfrom the group of: xanthan lyases, proteases, amylases, lichenases,lipases, cutinases, cellulases, endoglucanases, xyloglucanases,pectinases, pectin lyases, xanthanases, peroxidases, haloperoxygenases,catalases and mannanases, or any mixture thereof.

In one embodiment, the present disclosure relates to a detergentcomposition comprising a composition comprising at least one (e.g. about1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) endoglucanase variant as describedherein, wherein said composition further comprises one or more detergentcomponents and one or more additional enzymes selected from the groupof: xanthan lyases, proteases, amylases, lichenases, lipases, cutinases,cellulases, endoglucanases, xyloglucanases, pectinases, pectin lyases,xanthanases, peroxidases, haloperoxygenases, catalases and mannanases,or any mixture thereof, preferably said detergent component comprises achelator; further preferably said chelator is EDTA or citrate.

In one embodiment, the present disclosure relates to a detergentcomposition comprising a composition comprising at least one (e.g. about1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) endoglucanase variant as describedherein, wherein said composition further comprises one or more detergentcomponents, wherein said detergent composition is in form of a bar, ahomogenous tablet, a tablet having two or more layers, a pouch havingone or more compartments, a regular or compact powder, a granule, apaste, a gel, or a regular, compact or concentrated liquid.

In one embodiment, the present disclosure relates to use of a detergentcomposition of the disclosure, wherein said use is selected from thegroup of: use for degrading xanthan gum and use in a cleaning process,such as laundry or hard surface cleaning such as dish wash.

In one embodiment, the present disclosure relates to use of a detergentcomposition of the invention, wherein said composition has an enzymedetergency benefit

In one embodiment the present disclosure relates to a method fordegrading xanthan gum comprising: applying a detergent composition ofthe disclosure to a xanthan gum.

In one embodiment the present disclosure relates to a method fordegrading xanthan gum comprising: applying a detergent composition ofthe disclosure to a xanthan gum, wherein said xanthan gum is on thesurface of a textile or hard surface, such as dish wash.

Polynucleotides

Disclosed herein are isolated polynucleotides encoding a variant of thepresent disclosure. Said isolated polynucleotides encoding a variant maycomprise an alteration in one or more positions in a chelator-inducedinstability region selected from the group of: i) region 1 correspondingto amino acids 95 to 105 of SEQ ID NO: 2, ii) region 2 corresponding toamino acids 115 to 138 of SEQ ID NO: 2, iii) region 3 corresponding toamino acids 210 to 251 of SEQ ID NO: 2, iv) region 4 corresponding toamino acids 267 to 301 of SEQ ID NO: 2, v) region 5 corresponding toamino acids 339 to 361 of SEQ ID NO: 2, vi) region 6 corresponding toamino acids 547 to 595 of SEQ ID NO: 2, vii) region 7 corresponding toamino acids 612 to 660 of SEQ ID NO: 2, viii) region 8 corresponding toamino acids 806 to 828 of SEQ ID NO: 2, and ix) region 9 correspondingto amino acids 839 to 1042 of SEQ ID NO: 2, wherein said variant has atleast about 60% and less than about 100% sequence identity to SEQ ID NO:2.

The techniques used to isolate or clone a polynucleotide are known inthe art and include isolation from genomic DNA or cDNA, or a combinationthereof. The cloning of the polynucleotides from genomic DNA can beeffected, e.g., by using the well-known polymerase chain reaction (PCR)or antibody screening of expression libraries to detect cloned DNAfragments with shared structural features. See, e.g., Innis et al.,1990, PCR: A Guide to Methods and Application, Academic Press, New York.Other nucleic acid amplification procedures such as ligase chainreaction (LCR), ligation activated transcription (LAT) andpolynucleotide-based amplification (NASBA) may be used. Thepolynucleotides may be cloned in a strain of Bacillus subtilis or E.coli, or a related organism and thus, for example, may be an allelic orspecies variant of the polypeptide encoding region of thepolynucleotide.

Modification of a polynucleotide encoding a polypeptide as describedherein may be necessary for synthesizing polypeptides substantiallysimilar to the polypeptide. The term “substantially similar” to thepolypeptide refers to non-naturally occurring forms of the polypeptide.These polypeptides may differ in some engineered way from thepolypeptide isolated from its native source, e.g., variants that differin specific activity, thermostability, pH optimum, or the like. Thevariants may be constructed on the basis of the polynucleotide presentedas the mature polypeptide coding sequence of SEQ ID NO: 1, e.g., asubsequence thereof, and/or by introduction of nucleotide substitutionsthat do not result in a change in the amino acid sequence of thepolypeptide, but which correspond to the codon usage of the hostorganism intended for production of the enzyme, or by introduction ofnucleotide substitutions that may give rise to a different amino acidsequence. For a general description of nucleotide substitution, see,Ford et al., (1991), ‘Protein Expression and Purification’, 2: 95-107.

Nucleic Acid Constructs

Also disclosed herein are nucleic acid constructs comprising apolynucleotide encoding a variant as described herein operably linked toone or more control sequences that direct the expression of the codingsequence in a suitable host cell under conditions compatible with thecontrol sequences. Thus, disclosed are also nucleic acid constructscomprising a polynucleotide encoding a variant comprising an alterationin one or more positions in a chelator-induced instability regionselected from the group of: i) region 1 corresponding to amino acids 95to 105 of SEQ ID NO: 2, ii) region 2 corresponding to amino acids 115 to138 of SEQ ID NO: 2, iii) region 3 corresponding to amino acids 210 to251 of SEQ ID NO: 2, iv) region 4 corresponding to amino acids 267 to301 of SEQ ID NO: 2, v) region 5 corresponding to amino acids 339 to 361of SEQ ID NO: 2, vi) region 6 corresponding to amino acids 547 to 595 ofSEQ ID NO: 2, vii) region 7 corresponding to amino acids 612 to 660 ofSEQ ID NO: 2, viii) region 8 corresponding to amino acids 806 to 828 ofSEQ ID NO: 2, and ix) region 9 corresponding to amino acids 839 to 1042of SEQ ID NO: 2, wherein said variant has at least about 60% and lessthan about 100% sequence identity to SEQ ID NO: 2 operably linked to oneor more control sequences that direct the expression of the codingsequence in a suitable host cell under conditions compatible with thecontrol sequences.

A polynucleotide may be manipulated in a variety of ways to provide forexpression of the polypeptide. Manipulation of the polynucleotide priorto its insertion into a vector may be desirable or necessary dependingon the expression vector. The techniques for modifying polynucleotidesutilizing recombinant DNA methods are well known in the art.

The control sequence may be a promoter, a polynucleotide that isrecognized by a host cell for expression of a polynucleotide encoding apolypeptide of the present disclosure. The promoter containstranscriptional control sequences that mediate the expression of thepolypeptide. The promoter may be any polynucleotide that showstranscriptional activity in the host cell including mutant, truncated,and hybrid promoters, and may be obtained from genes encodingextracellular or intracellular polypeptides either homologous orheterologous to the host cell.

Examples of suitable promoters for directing transcription of thenucleic acid constructs of the present disclosure in a bacterial hostcell are the promoters obtained from the Bacillus amyloliquefaciensalpha-amylase gene (amyQ), Bacillus licheniformis alpha-amylase gene(amyL), Bacillus licheniformis penicillinase gene (penP), Bacillusstearothermophilus maltogenic amylase gene (amyM), Bacillus subtilislevansucrase gene (sacB), Bacillus subtilis xylA and xylB genes,Bacillus thuringiensis cryIIIA gene (Agaisse and Lereclus, 1994,Molecular Microbiology 13: 97-107), E. coli lac operon, E. coli trcpromoter (Egon et al., 1988, Gene 69: 301-315), Streptomyces coelicoloragarase gene (dagA), and prokaryotic beta-lactamase gene (Villa-Kamaroffet al., 1978, Proc. Natl. Acad. Sci. USA 75: 3727-3731), as well as thetac promoter (DeBoer et al., 1983, Proc. Natl. Acad. Sci. USA 80:21-25). Further promoters are described in “Useful proteins fromrecombinant bacteria” in Gilbert et al., 1980, Scientific American 242:74-94; and in Sambrook et al., 1989, supra. Examples of tandem promotersare disclosed in WO 99/43835.

Examples of suitable promoters for directing transcription of thenucleic acid constructs of the present disclosure in a filamentousfungal host cell are promoters obtained from the genes for Aspergillusnidulans acetamidase, Aspergillus niger neutral alpha-amylase,Aspergillus niger acid stable alpha-amylase, Aspergillus niger orAspergillus awamori glucoamylase (glaA), Aspergillus oryzae TAKAamylase, Aspergillus oryzae alkaline protease, Aspergillus oryzae triosephosphate isomerase, Fusarium oxysporum trypsin-like protease (WO96/00787), Fusarium venenatum amyloglucosidase (WO 00/56900), Fusariumvenenatum Daria (WO 00/56900), Fusarium venenatum Quinn (WO 00/56900),Rhizomucor miehei lipase, Rhizomucor miehei aspartic proteinase,Trichoderma reesei beta-glucosidase, Trichoderma reeseicellobiohydrolase I, Trichoderma reesei cellobiohydrolase II,Trichoderma reesei endoglucanase I, Trichoderma reesei endoglucanase II,Trichoderma reesei endoglucanase III, Trichoderma reesei endoglucanaseIV, Trichoderma reesei endoglucanase V, Trichoderma reesei xylanase I,Trichoderma reesei xylanase II, Trichoderma reesei beta-xylosidase, aswell as the NA2-tpi promoter (a modified promoter from an Aspergillusneutral alpha-amylase gene in which the untranslated leader has beenreplaced by an untranslated leader from an Aspergillus triose phosphateisomerase gene; non-limiting examples include modified promoters from anAspergillus niger neutral alpha-amylase gene in which the untranslatedleader has been replaced by an untranslated leader from an Aspergillusnidulans or Aspergillus oryzae triose phosphate isomerase gene); andmutant, truncated, and hybrid promoters thereof.

In a yeast host, useful promoters are obtained from the genes forSaccharomyces cerevisiae enolase (ENO-1), Saccharomyces cerevisiaegalactokinase (GAL1), Saccharomyces cerevisiae alcoholdehydrogenase/glyceraldehyde-3-phosphate dehydrogenase (ADH1, ADH2/GAP),Saccharomyces cerevisiae triose phosphate isomerase (TPI), Saccharomycescerevisiae metallothionein (CUP1), and Saccharomyces cerevisiae3-phosphoglycerate kinase. Other useful promoters for yeast host cellsare described by Romanos et al., 1992, Yeast 8: 423-488.

The control sequence may also be a transcription terminator, which isrecognized by a host cell to terminate transcription. The terminator isoperably linked to the 3′-terminus of the polynucleotide encoding thepolypeptide. Any terminator that is functional in the host cell may beused in the present disclosure.

Preferred terminators for bacterial host cells are obtained from thegenes for Bacillus clausii alkaline protease (aprH), Bacilluslicheniformis alpha-amylase (amyL), and Escherichia coli ribosomal RNA(rrnB).

Preferred terminators for filamentous fungal host cells are obtainedfrom the genes for Aspergillus nidulans anthranilate synthase,Aspergillus niger glucoamylase, Aspergillus niger alpha-glucosidase,Aspergillus oryzae TAKA amylase, and Fusarium oxysporum trypsin-likeprotease.

Preferred terminators for yeast host cells are obtained from the genesfor Saccharomyces cerevisiae enolase, Saccharomyces cerevisiaecytochrome C (CYC1), and Saccharomyces cerevisiaeglyceraldehyde-3-phosphate dehydrogenase. Other useful terminators foryeast host cells are described by Romanos et al., 1992, supra.

The control sequence may also be an mRNA stabilizer region downstream ofa promoter and upstream of the coding sequence of a gene which increasesexpression of the gene.

Examples of suitable mRNA stabilizer regions are obtained from aBacillus thuringiensis cryIIIA gene (WO 94/25612) and a Bacillussubtilis SP82 gene (Hue et al., 1995, Journal of Bacteriology 177:3465-3471).

The control sequence may also be a leader, a nontranslated region of anmRNA that is important for translation by the host cell. The leader isoperably linked to the 5′-terminus of the polynucleotide encoding thepolypeptide. Any leader that is functional in the host cell may be used.

Preferred leaders for filamentous fungal host cells are obtained fromthe genes for Aspergillus oryzae TAKA amylase and Aspergillus nidulanstriose phosphate isomerase.

Suitable leaders for yeast host cells are obtained from the genes forSaccharomyces cerevisiae enolase (ENO-1), Saccharomyces cerevisiae3-phosphoglycerate kinase, Saccharomyces cerevisiae alpha-factor, andSaccharomyces cerevisiae alcoholdehydrogenase/glyceraldehyde-3-phosphate dehydrogenase (ADH2/GAP).

The control sequence may also be a polyadenylation sequence, a sequenceoperably linked to the 3′-terminus of the polynucleotide and, whentranscribed, is recognized by the host cell as a signal to addpolyadenosine residues to transcribed mRNA. Any polyadenylation sequencethat is functional in the host cell may be used.

Preferred polyadenylation sequences for filamentous fungal host cellsare obtained from the genes for Aspergillus nidulans anthranilatesynthase, Aspergillus niger glucoamylase, Aspergillus nigeralpha-glucosidase Aspergillus oryzae TAKA amylase, and Fusariumoxysporum trypsin-like protease.

Useful polyadenylation sequences for yeast host cells are described byGuo and Sherman, 1995, Mol. Cellular Biol. 15: 5983-5990.

The control sequence may also be a signal peptide coding region thatencodes a signal peptide linked to the N-terminus of a polypeptide anddirects the polypeptide into the cell's secretory pathway. The 5′-end ofthe coding sequence of the polynucleotide may inherently contain asignal peptide coding sequence naturally linked in translation readingframe with the segment of the coding sequence that encodes thepolypeptide. Alternatively, the 5′-end of the coding sequence maycontain a signal peptide coding sequence that is foreign to the codingsequence. A foreign signal peptide coding sequence may be required wherethe coding sequence does not naturally contain a signal peptide codingsequence. Alternatively, a foreign signal peptide coding sequence maysimply replace the natural signal peptide coding sequence in order toenhance secretion of the polypeptide. However, any signal peptide codingsequence that directs the expressed polypeptide into the secretorypathway of a host cell may be used.

Effective signal peptide coding sequences for bacterial host cells arethe signal peptide coding sequences obtained from the genes for BacillusNCIB 11837 maltogenic amylase, Bacillus licheniformis subtilisin,Bacillus licheniformis beta-lactamase, Bacillus stearothermophilusalpha-amylase, Bacillus stearothermophilus neutral proteases (nprT,nprS, nprM), and Bacillus subtilis prsA. Further signal peptides aredescribed by Simonen and Palva, 1993, Microbiological Reviews 57:109-137.

Effective signal peptide coding sequences for filamentous fungal hostcells are the signal peptide coding sequences obtained from the genesfor Aspergillus niger neutral amylase, Aspergillus niger glucoamylase,Aspergillus oryzae TAKA amylase, Humicola insolens cellulase, Humicolainsolens endoglucanase V, Humicola lanuginosa lipase, and Rhizomucormiehei aspartic proteinase.

Useful signal peptides for yeast host cells are obtained from the genesfor Saccharomyces cerevisiae alpha-factor and Saccharomyces cerevisiaeinvertase. Other useful signal peptide coding sequences are described byRomanos et al., 1992, supra.

The control sequence may also be a propeptide coding sequence thatencodes a propeptide positioned at the N-terminus of a polypeptide. Theresultant polypeptide is known as a proenzyme or propolypeptide (or azymogen in some cases). A propolypeptide is generally inactive and canbe converted to an active polypeptide by catalytic or autocatalyticcleavage of the propeptide from the propolypeptide. The propeptidecoding sequence may be obtained from the genes for Bacillus subtilisalkaline protease (aprE), Bacillus subtilis neutral protease (nprT),Myceliophthora thermophila laccase (WO 95/33836), Rhizomucor mieheiaspartic proteinase, and Saccharomyces cerevisiae alpha-factor.

Where both signal peptide and propeptide sequences are present, thepropeptide sequence is positioned next to the N-terminus of apolypeptide and the signal peptide sequence is positioned next to theN-terminus of the propeptide sequence.

It may also be desirable to add regulatory sequences that regulateexpression of the polypeptide relative to the growth of the host cell.Examples of regulatory systems are those that cause expression of thegene to be turned on or off in response to a chemical or physicalstimulus, including the presence of a regulatory compound. Regulatorysystems in prokaryotic systems include the lac, tac, and trp operatorsystems. In yeast, the ADH2 system or GAL1 system may be used. Infilamentous fungi, the Aspergillus niger glucoamylase promoter,Aspergillus oryzae TAKA alpha-amylase promoter, and Aspergillus oryzaeglucoamylase promoter may be used. Other examples of regulatorysequences are those that allow for gene amplification. In eukaryoticsystems, these regulatory sequences include the dihydrofolate reductasegene that is amplified in the presence of methotrexate, and themetallothionein genes that are amplified with heavy metals. In thesecases, the polynucleotide encoding the polypeptide would be operablylinked with the regulatory sequence.

Expression Vectors

Also disclosed herein are recombinant expression vectors comprising apolynucleotide encoding a variant as described herein, a promoter, andtranscriptional and translational stop signals. Thus, described hereinare recombinant expression vectors comprising a polynucleotide encodinga variant comprising an alteration in one or more positions in achelator-induced instability region selected from the group of: i)region 1 corresponding to amino acids 95 to 105 of SEQ ID NO: 2, ii)region 2 corresponding to amino acids 115 to 138 of SEQ ID NO: 2, iii)region 3 corresponding to amino acids 210 to 251 of SEQ ID NO: 2, iv)region 4 corresponding to amino acids 267 to 301 of SEQ ID NO: 2, v)region 5 corresponding to amino acids 339 to 361 of SEQ ID NO: 2, vi)region 6 corresponding to amino acids 547 to 595 of SEQ ID NO: 2, vii)region 7 corresponding to amino acids 612 to 660 of SEQ ID NO: 2, viii)region 8 corresponding to amino acids 806 to 828 of SEQ ID NO: 2, andix) region 9 corresponding to amino acids 839 to 1042 of SEQ ID NO: 2,wherein said variant has at least about 60% and less than about 100%sequence identity to SEQ ID NO: 2, a promotor, and transcriptional andtranslational stop signals.

The various nucleotide and control sequences may be joined together toproduce a recombinant expression vector that may include one or moreconvenient restriction sites to allow for insertion or substitution ofthe polynucleotide encoding the polypeptide at such sites.Alternatively, the polynucleotide may be expressed by inserting thepolynucleotide or a nucleic acid construct comprising the polynucleotideinto an appropriate vector for expression. In creating the expressionvector, the coding sequence is located in the vector so that the codingsequence is operably linked with the appropriate control sequences forexpression.

The recombinant expression vector may be any vector (e.g., a plasmid orvirus) that can be conveniently subjected to recombinant DNA proceduresand can bring about expression of the polynucleotide. The choice of thevector will typically depend on the compatibility of the vector with thehost cell into which the vector is to be introduced. The vector may be alinear or closed circular plasmid.

The vector may be an autonomously replicating vector, i.e., a vectorthat exists as an extrachromosomal entity, the replication of which isindependent of chromosomal replication, e.g., a plasmid, anextrachromosomal element, a minichromosome, or an artificial chromosome.The vector may contain any mechanism for assuring self-replication.Alternatively, the vector may be one that, when introduced into the hostcell, is integrated into the genome and replicated together with thechromosome(s) into which it has been integrated. Furthermore, a singlevector or plasmid or two or more vectors or plasmids that togethercontain the total DNA to be introduced into the genome of the host cell,or a transposon, may be used.

The vector preferably contains one or more selectable markers thatpermit easy selection of transformed, transfected, transduced, or thelike cells. A selectable marker is a gene the product of which providesfor biocide or viral resistance, resistance to heavy metals, prototrophyto auxotrophs, and the like.

Examples of bacterial selectable markers are Bacillus licheniformis orBacillus subtilis dal genes, or markers that confer antibioticresistance such as ampicillin, chloramphenicol, kanamycin, neomycin,spectinomycin, or tetracycline resistance. Suitable markers for yeasthost cells include, but are not limited to, ADE2, HIS3, LEU2, LYS2,MET3, TRP1, and URA3. Selectable markers for use in a filamentous fungalhost cell include, but are not limited to, amdS (acetamidase), argB(ornithine carbamoyltransferase), bar (phosphinothricinacetyltransferase), hph (hygromycin phosphotransferase), niaD (nitratereductase), pyrG (orotidine-5′-phosphate decarboxylase), sC (sulfateadenyltransferase), and trpC (anthranilate synthase), as well asequivalents thereof. Preferred for use in an Aspergillus cell areAspergillus nidulans or Aspergillus oryzae amdS and pyrG genes and aStreptomyces hygroscopicus bar gene.

The vector preferably contains an element(s) that permits integration ofthe vector into the host cell's genome or autonomous replication of thevector in the cell independent of the genome.

For integration into the host cell genome, the vector may rely on thepolynucleotide's sequence encoding the polypeptide or any other elementof the vector for integration into the genome by homologous ornon-homologous recombination. Alternatively, the vector may containadditional polynucleotides for directing integration by homologousrecombination into the genome of the host cell at a precise location(s)in the chromosome(s). To increase the likelihood of integration at aprecise location, the integrational elements should contain a sufficientnumber of nucleic acids, such as about 100 to about 10,000 base pairs,about 400 to about 10,000 base pairs, and about 800 to about 10,000 basepairs, which have a high degree of sequence identity to thecorresponding target sequence to enhance the probability of homologousrecombination. The integrational elements may be any sequence that ishomologous with the target sequence in the genome of the host cell.Furthermore, the integrational elements may be non-encoding or encodingpolynucleotides. On the other hand, the vector may be integrated intothe genome of the host cell by non-homologous recombination.

For autonomous replication, the vector may further comprise an origin ofreplication enabling the vector to replicate autonomously in the hostcell in question. The origin of replication may be any plasmidreplicator mediating autonomous replication that functions in a cell.The term “origin of replication” or “plasmid replicator” means apolynucleotide that enables a plasmid or vector to replicate in vivo.

Examples of bacterial origins of replication are the origins ofreplication of plasmids pBR322, pUC19, pACYC177, and pACYC184 permittingreplication in E. coli, and pUB110, pE194, pTA1060, and pAMβ1 permittingreplication in Bacillus.

Examples of origins of replication for use in a yeast host cell are the2 micron origin of replication, ARS1, ARS4, the combination of ARS1 andCEN3, and the combination of ARS4 and CEN6.

Examples of origins of replication useful in a filamentous fungal cellare AMA1 and ANS1 (Gems et al., 1991, Gene 98: 61-67; Cullen et al.,1987, Nucleic Acids Res. 15: 9163-9175; WO 00/24883). Isolation of theAMA1 gene and construction of plasmids or vectors comprising the genecan be accomplished according to the methods disclosed in WO 00/24883.

More than one copy of a polynucleotide of the present disclosure may beinserted into a host cell to increase production of a polypeptide. Anincrease in the copy number of the polynucleotide can be obtained byintegrating at least one additional copy of the sequence into the hostcell genome or by including an amplifiable selectable marker gene withthe polynucleotide where cells containing amplified copies of theselectable marker gene, and thereby additional copies of thepolynucleotide, can be selected for by cultivating the cells in thepresence of the appropriate selectable agent.

The procedures used to ligate the elements described above to constructthe recombinant expression vectors of the present disclosure are wellknown to one skilled in the art (see, e.g., Sambrook et al., 1989,supra).

Host Cells

Further disclosed are recombinant host cells, comprising apolynucleotide encoding a variant as described herein operably linked toone or more control sequences that direct the production of a variantdescribed herein. Thus, described are also recombinant host cells,comprising a polynucleotide encoding a variant comprising an alterationin one or more positions in a chelator-induced instability regionselected from the group of: i) region 1 corresponding to amino acids 95to 105 of SEQ ID NO: 2, ii) region 2 corresponding to amino acids 115 to138 of SEQ ID NO: 2, iii) region 3 corresponding to amino acids 210 to251 of SEQ ID NO: 2, iv) region 4 corresponding to amino acids 267 to301 of SEQ ID NO: 2, v) region 5 corresponding to amino acids 339 to 361of SEQ ID NO: 2, vi) region 6 corresponding to amino acids 547 to 595 ofSEQ ID NO: 2, vii) region 7 corresponding to amino acids 612 to 660 ofSEQ ID NO: 2, viii) region 8 corresponding to amino acids 806 to 828 ofSEQ ID NO: 2, and ix) region 9 corresponding to amino acids 839 to 1042of SEQ ID NO: 2, wherein said variant has at least about 60% and lessthan about 100% sequence identity to SEQ ID NO: 2 operably linked to oneor more control sequences that direct the production of a variantcomprising an alteration in one or more positions in a chelator-inducedinstability region selected from the group of: i) region 1 correspondingto amino acids 95 to 105 of SEQ ID NO: 2, ii) region 2 corresponding toamino acids 115 to 138 of SEQ ID NO: 2, iii) region 3 corresponding toamino acids 210 to 251 of SEQ ID NO: 2, iv) region 4 corresponding toamino acids 267 to 301 of SEQ ID NO: 2, v) region 5 corresponding toamino acids 339 to 361 of SEQ ID NO: 2, vi) region 6 corresponding toamino acids 547 to 595 of SEQ ID NO: 2, vii) region 7 corresponding toamino acids 612 to 660 of SEQ ID NO: 2, viii) region 8 corresponding toamino acids 806 to 828 of SEQ ID NO: 2, and ix) region 9 correspondingto amino acids 839 to 1042 of SEQ ID NO: 2, wherein said variant has atleast about 60% and less than about 100% sequence identity to SEQ ID NO:2.

A construct or vector comprising a polynucleotide is introduced into ahost cell so that the construct or vector is maintained as a chromosomalintegrant or as a self-replicating extra-chromosomal vector as describedearlier. The term “host cell” encompasses any progeny of a parent cellthat is not identical to the parent cell due to mutations that occurduring replication. The choice of a host cell will to a large extentdepend upon the gene encoding the polypeptide and its source.

The host cell may be any cell useful in the recombinant production of apolypeptide of the present disclosure, e.g., a prokaryote or aeukaryote.

The prokaryotic host cell may be any Gram-positive or Gram-negativebacterium. Gram-positive bacteria include, but are not limited to,Bacillus, Clostridium, Enterococcus, Geobacillus, Lactobacillus,Lactococcus, Oceanobacillus, Staphylococcus, Streptococcus, andStreptomyces. Gram-negative bacteria include, but are not limited to,Campylobacter, E. coli, Flavobacterium, Fusobacterium, Helicobacter,Ilyobacter, Neisseria, Pseudomonas, Salmonella, and Ureaplasma.

The bacterial host cell may be any Bacillus cell including, but notlimited to, Bacillus alkalophilus, Bacillus amyloliquefaciens, Bacillusbrevis, Bacillus circulars, Bacillus clausii, Bacillus coagulans,Bacillus firmus, Bacillus lautus, Bacillus lentus, Bacilluslicheniformis, Bacillus megaterium, Bacillus pumilus, Bacillusstearothermophilus, Bacillus subtilis, and Bacillus thuringiensis cells.

The bacterial host cell may also be any Streptococcus cell including,but not limited to, Streptococcus equisimilis, Streptococcus pyogenes,Streptococcus uberis, and Streptococcus equi subsp. Zooepidemicus cells.

The bacterial host cell may also be any Streptomyces cell including, butnot limited to, Streptomyces achromogenes, Streptomyces avermitilis,Streptomyces coelicolor, Streptomyces griseus, and Streptomyces lividanscells.

The introduction of DNA into a Bacillus cell may be effected byprotoplast transformation (see, e.g., Chang and Cohen, 1979, Mol. Gen.Genet. 168: 111-115), competent cell transformation (see, e.g., Youngand Spizizen, 1961, J. Bacteriol. 81: 823-829, or Dubnau andDavidoff-Abelson, 1971, J. Mol. Biol. 56: 209-221), electroporation(see, e.g., Shigekawa and Dower, 1988, Biotechniques 6: 742-751), orconjugation (see, e.g., Koehler and Thorne, 1987, J. Bacteriol. 169:5271-5278). The introduction of DNA into an E. coli cell may be effectedby protoplast transformation (see, e.g., Hanahan, 1983, J. Mol. Biol.166: 557-580) or electroporation (see, e.g., Dower et al., 1988, NucleicAcids Res. 16: 6127-6145). The introduction of DNA into a Streptomycescell may be effected by protoplast transformation, electroporation (see,e.g., Gong et al., 2004, Folia Microbiol. (Praha) 49: 399-405),conjugation (see, e.g., Mazodier et al., 1989, J. Bacteriol. 171:3583-3585), or transduction (see, e.g., Burke et al., 2001, Proc. Natl.Acad. Sci. USA 98: 6289-6294). The introduction of DNA into aPseudomonas cell may be effected by electroporation (see, e.g., Choi etal., 2006, J. Microbiol. Methods 64: 391-397) or conjugation (see, e.g.,Pinedo and Smets, 2005, Appl. Environ. Microbiol. 71: 51-57). Theintroduction of DNA into a Streptococcus cell may be effected by naturalcompetence (see, e.g., Perry and Kuramitsu, 1981, Infect. Immun. 32:1295-1297), protoplast transformation (see, e.g., Catt and Jollick,1991, Microbios 68: 189-207), electroporation (see, e.g., Buckley etal., 1999, Appl. Environ. Microbiol. 65: 3800-3804), or conjugation(see, e.g., Clewell, 1981, Microbiol. Rev. 45: 409-436). However, anymethod known in the art for introducing DNA into a host cell can beused.

The host cell may also be a eukaryote, such as a mammalian, insect,plant, or fungal cell.

The host cell may be a fungal cell. “Fungi” as used herein includes thephyla Ascomycota, Basidiomycota, Chytridiomycota, and Zygomycota as wellas the Oomycota and all mitosporic fungi (as defined by Hawksworth etal., In, Ainsworth and Bisby's Dictionary of The Fungi, 8th edition,1995, CAB International, University Press, Cambridge, UK).

The fungal host cell may be a yeast cell. “Yeast” as used hereinincludes ascosporogenous yeast (Endomycetales), basidiosporogenousyeast, and yeast belonging to the Fungi Imperfecti (Blastomycetes).Since the classification of yeast may change in the future, for thepurposes of this disclosure, yeast shall be defined as described inBiology and Activities of Yeast (Skinner, Passmore, and Davenport,editors, Soc. App. Bacteriol. Symposium Series No. 9, 1980).

The yeast host cell may be a Candida, Hansenula, Kluyveromyces, Pichia,Saccharomyces, Schizosaccharomyces, or Yarrowia cell, such as aKluyveromyces lactis, Saccharomyces carlsbergensis, Saccharomycescerevisiae, Saccharomyces diastaticus, Saccharomyces douglasii,Saccharomyces kluyveri, Saccharomyces norbensis, Saccharomycesoviformis, or Yarrowia lipolytica cell.

The fungal host cell may be a filamentous fungal cell. “Filamentousfungi” include all filamentous forms of the subdivision Eumycota andOomycota (as defined by Hawksworth et al., 1995, supra). The filamentousfungi are generally exemplified by a mycelial wall composed of chitin,cellulose, glucan, chitosan, mannan, and other complex polysaccharides.Vegetative growth is by hyphal elongation and carbon catabolism isobligately aerobic. In contrast, vegetative growth by yeasts such asSaccharomyces cerevisiae is by budding of a unicellular thallus andcarbon catabolism may be fermentative.

The filamentous fungal host cell may be an Acremonium, Aspergillus,Aureobasidium, Bjerkandera, Ceriporiopsis, Chrysosporium, Coprinus,Coriolus, Cryptococcus, Filibasidium, Fusarium, Humicola, Magnaporthe,Mucor, Myceliophthora, Neocallimastix, Neurospora, Paecilomyces,Penicillium, Phanerochaete, Phlebia, Piromyces, Pleurotus,Schizophyllum, Talaromyces, Thermoascus, Thielavia, Tolypocladium,Trametes, or Trichoderma cell.

For example, the filamentous fungal host cell may be an Aspergillusawamori, Aspergillus foetidus, Aspergillus fumigatus, Aspergillusjaponicus, Aspergillus nidulans, Aspergillus niger, Aspergillus oryzae,Bjerkandera adusta, Ceriporiopsis aneirina, Ceriporiopsis caregiea,Ceriporiopsis gilvescens, Ceriporiopsis pannocinta, Ceriporiopsisrivulosa, Ceriporiopsis subrufa, Ceriporiopsis subvermispora,Chrysosporium inops, Chrysosporium keratinophilum, Chrysosporiumlucknowense, Chrysosporium merdarium, Chrysosporium pannicola,Chrysosporium queenslandicum, Chrysosporium tropicum, Chrysosporiumzonatum, Coprinus cinereus, Coriolus hirsutus, Fusarium bactridioides,Fusarium cerealis, Fusarium crookwellense, Fusarium culmorum, Fusariumgraminearum, Fusarium graminum, Fusarium heterosporum, Fusarium negundi,Fusarium oxysporum, Fusarium reticulatum, Fusarium roseum, Fusariumsambucinum, Fusarium sarcochroum, Fusarium sporotrichioides, Fusariumsulphureum, Fusarium torulosum, Fusarium trichothecioides, Fusariumvenenatum, Humicola insolens, Humicola lanuginosa, Mucor miehei,Myceliophthora thermophila, Neurospora crassa, Penicillium purpurogenum,Phanerochaete chrysosporium, Phlebia radiata, Pleurotus eryngii,Thielavia terrestris, Trametes villosa, Trametes versicolor, Trichodermaharzianum, Trichoderma koningii, Trichoderma longibrachiatum,Trichoderma reesei, or Trichoderma viride cell.

Fungal cells may be transformed by a process involving protoplastformation, transformation of the protoplasts, and regeneration of thecell wall in a manner known per se. Suitable procedures fortransformation of Aspergillus and Trichoderma host cells are describedin EP 238023, Yelton et al., 1984, Proc. Natl. Acad. Sci. USA 81:1470-1474, and Christensen et al., 1988, Bio/Technology 6: 1419-1422.Suitable methods for transforming Fusarium species are described byMalardier et al., 1989, Gene 78: 147-156, and WO 96/00787. Yeast may betransformed using the procedures described by Becker and Guarente, InAbelson, J. N. and Simon, M. I., editors, Guide to Yeast Genetics andMolecular Biology, Methods in Enzymology, Volume 194, pp 182-187,Academic Press, Inc., New York; Ito et al., 1983, J. Bacteriol. 153:163; and Hinnen et al., 1978, Proc. Natl. Acad. Sci. USA 75: 1920.

Methods of Production

Also described are methods of producing (e.g., in vitro or ex vivomethods) a variant, comprising: (a) cultivating a host cell as describedabove under conditions suitable for expression of the variant; and (b)recovering the variant. Thus, disclosed are also methods of producing(e.g., in vitro or ex vivo methods) a variant, comprising: (a)cultivating a host cell comprising a polynucleotide encoding a variantcomprising an alteration in one or more positions in a chelator-inducedinstability region selected from the group of: i) region 1 correspondingto amino acids 95 to 105 of SEQ ID NO: 2, ii) region 2 corresponding toamino acids 115 to 138 of SEQ ID NO: 2, iii) region 3 corresponding toamino acids 210 to 251 of SEQ ID NO: 2, iv) region 4 corresponding toamino acids 267 to 301 of SEQ ID NO: 2, v) region 5 corresponding toamino acids 339 to 361 of SEQ ID NO: 2, vi) region 6 corresponding toamino acids 547 to 595 of SEQ ID NO: 2, vii) region 7 corresponding toamino acids 612 to 660 of SEQ ID NO: 2, viii) region 8 corresponding toamino acids 806 to 828 of SEQ ID NO: 2, and ix) region 9 correspondingto amino acids 839 to 1042 of SEQ ID NO: 2, wherein said variant has atleast about 60% and less than about 100% sequence identity to SEQ ID NO:2, under conditions suitable for expression of the variant, and (b)optionally, recovering the variant.

Further disclosed are methods of producing (e.g., in vitro or ex vivomethods) a variant s described herein, comprising (a) cultivating acell, which in its wild-type form produces the polypeptide, underconditions conducive for production of the polypeptide; and (b)recovering the polypeptide. In a preferred aspect, the cell is aPaenibacillus cell, or a Microbacterium cell.

Also disclosed are methods of producing (e.g., in vitro or ex vivomethods) a variant as described herein, comprising (a) cultivating arecombinant host cell of the present disclosure under conditionsconducive for production of the polypeptide; and (b) recovering thepolypeptide.

The host cells are cultivated in a nutrient medium suitable forproduction of the polypeptide using methods known in the art. Forexample, the cell may be cultivated by shake flask cultivation, orsmall-scale or large-scale fermentation (including continuous, batch,fed-batch, or solid state fermentations) in laboratory or industrialfermentors performed in a suitable medium and under conditions allowingthe polypeptide to be expressed and/or isolated. The cultivation takesplace in a suitable nutrient medium comprising carbon and nitrogensources and inorganic salts, using procedures known in the art. Suitablemedia are available from commercial suppliers or may be preparedaccording to published compositions (e.g., in catalogues of the AmericanType Culture Collection). If the polypeptide is secreted into thenutrient medium, the polypeptide can be recovered directly from themedium. If the polypeptide is not secreted, it can be recovered fromcell lysates.

The variant polypeptide may be detected using methods known in the artthat are specific for the polypeptides such as methods for determiningcellulose or xanthan lyase activity. These detection methods include,but are not limited to, use of specific antibodies, formation of anenzyme product, or disappearance of an enzyme substrate. For example, anenzyme assay may be used to determine the activity of the polypeptide.

The variant polypeptide may be recovered using methods known in the art.For example, the polypeptide may be recovered from the nutrient mediumby conventional procedures including, but not limited to, collection,centrifugation, filtration, extraction, spray-drying, evaporation, orprecipitation.

The variant polypeptide may be purified by a variety of procedures knownin the art including, but not limited to, chromatography (e.g., ionexchange, affinity, hydrophobic, chromatofocusing, and size exclusion),electrophoretic procedures (e.g., preparative isoelectric focusing),differential solubility (e.g., ammonium sulfate precipitation),SDS-PAGE, or extraction (see, e.g., Protein Purification, Janson andRyden, editors, VCH Publishers, New York, 1989) to obtain substantiallypure polypeptides.

In an alternative aspect, the variant polypeptide is not recovered, butrather a host cell expressing the polypeptide is used as a source of thevariant polypeptide.

Compositions

In one certain aspect, the variants as contemplated herein have improvedstability in detergents compared to a parent enzyme or compared to anendoglucanase having the identical amino acid sequence of the variant,but not having an alteration (e.g., a substitution, deletion orinsertion) at one or more of the specified positions or compared to theendoglucanase with SEQ ID NO: 2, wherein activity and/or stability indetergent is measured as disclosed in example 4 herein. Thus, in oneembodiment, the present disclosure relates to detergent compositionscomprising a variant comprising an alteration in one or more positionsin a chelator-induced instability region selected from the group of: i)region 1 corresponding to amino acids 95 to 105 of SEQ ID NO: 2, ii)region 2 corresponding to amino acids 115 to 138 of SEQ ID NO: 2, iii)region 3 corresponding to amino acids 210 to 251 of SEQ ID NO: 2, iv)region 4 corresponding to amino acids 267 to 301 of SEQ ID NO: 2, v)region 5 corresponding to amino acids 339 to 361 of SEQ ID NO: 2, vi)region 6 corresponding to amino acids 547 to 595 of SEQ ID NO: 2, vii)region 7 corresponding to amino acids 612 to 660 of SEQ ID NO: 2, viii)region 8 corresponding to amino acids 806 to 828 of SEQ ID NO: 2, andix) region 9 corresponding to amino acids 839 to 1042 of SEQ ID NO: 2,wherein said variant has at least about 60% and less than about 100%sequence identity to SEQ ID NO: 2, wherein the variant has an improvedstability in the detergent composition.

Besides enzymes the detergent compositions may comprise additionalcomponents. The choice of additional components is within the skill ofthe artisan and includes conventional ingredients, including theexemplary non-limiting components set forth below. The choice ofcomponents may include, for fabric care, the consideration of the typeof fabric to be cleaned, the type and/or degree of soiling, thetemperature at which cleaning is to take place, and the formulation ofthe detergent product. Although components mentioned below arecategorized by general header according to a particular functionality,this is not to be construed as a limitation, as a component may compriseadditional functionalities as will be appreciated by the skilledartisan.

The detergent composition may be suitable for the laundring of textilessuch as e.g. fabrics, cloths or linen, or for cleaning hard surfacessuch as e.g. floors, tables, or dish wash.

Detergent Compositions

In one embodiment, a variant as described herein may be added to adetergent composition in an amount corresponding to from about 0.0001 toabout 200 mg of enzyme protein, such as from about 0.0005 to about 100mg of enzyme protein, preferably from about 0.001 to about 30 mg ofenzyme protein, more preferably from about 0.005 to about 8 mg of enzymeprotein, even more preferably from about 0.01 to about 2 mg of enzymeprotein per litre of wash liquor.

A composition for use in automatic dishwash (ADW), for example, mayinclude from about 0.0001% to about 50%, such as from about 0.001% toabout 20%, such as from about 0.01% to about 10%, such as from about0.05 to about 5% of enzyme protein by weight of the composition.

A composition for use in laundry granulation or a solid/granular laundrycomposition in general, for example, may include from about 0.0001% toabout 50%, such as from about 0.001% to about 20%, such as from about0.01% to about 10%, such as from about 0.05% to about 5% of enzymeprotein by weight of the composition.

A composition for use in laundry liquid, for example, may include fromabout 0.0001% to about 10%, such as from about 0.001 to about 7%, suchas from about 0.1% to about 5% of enzyme protein by weight of thecomposition.

The enzyme(s) of the detergent composition of the disclosure may bestabilized using conventional stabilizing agents, e.g., a polyol such aspropylene glycol or glycerol, a sugar or sugar alcohol, lactic acid,boric acid, or a boric acid derivative, e.g., an aromatic borate ester,or a phenyl boronic acid derivative such as 4-formylphenyl boronic acid,and the composition may be formulated as described in, for example,WO92/19709 and WO92/19708.

In certain markets different wash conditions and, as such, differenttypes of detergents are used. This is disclosed in e.g. EP 1 025 240.For example, In Asia (Japan) a low detergent concentration system isused, while the United States uses a medium detergent concentrationsystem, and Europe uses a high detergent concentration system.

A low detergent concentration system includes detergents where less thanabout 800 ppm of detergent components are present in the wash water.Japanese detergents are typically considered low detergent concentrationsystem as they have approximately 667 ppm of detergent componentspresent in the wash water.

A medium detergent concentration includes detergents where between about800 ppm and about 2000 ppm of detergent components are present in thewash water. North American detergents are generally considered to bemedium detergent concentration systems as they have approximately 975ppm of detergent components present in the wash water.

A high detergent concentration system includes detergents where greaterthan about 2000 ppm of detergent components are present in the washwater. European detergents are generally considered to be high detergentconcentration systems as they have approximately 4500-5000 ppm ofdetergent components in the wash water.

Latin American detergents are generally high suds phosphate builderdetergents and the range of detergents used in Latin America can fall inboth the medium and high detergent concentrations as they range fromabout 1500 ppm to about 6000 ppm of detergent components in the washwater. Such detergent compositions are all embodiments of thedisclosure.

A polypeptide of the present disclosure may also be incorporated in thedetergent formulations disclosed in WO97/07202, which is herebyincorporated by reference.

Surfactants

The detergent composition may comprise one or more surfactants, whichmay be anionic and/or cationic and/or non-ionic and/or semi-polar and/orzwitterionic, or a mixture thereof. In a particular embodiment, thedetergent composition includes a mixture of one or more nonionicsurfactants and one or more anionic surfactants. The surfactant(s) istypically present at a level of from about 0.1% to about 60% by weight,such as about 1% to about 40%, or about 3% to about 20%, or about 3% toabout 10%. The surfactant(s) is chosen based on the desired cleaningapplication, and includes any conventional surfactant(s) known in theart. Any surfactant known in the art for use in detergents may beutilized.

When included therein the detergent will usually comprise from about 1%to about 40% by weight, such as from about 5% to about 30%, includingfrom about 5% to about 15%, or from about 20% to about 25% of an anionicsurfactant. Non-limiting examples of anionic surfactants includesulfates and sulfonates, in particular, linear alkylbenzenesulfonates(LAS), isomers of LAS, branched alkylbenzenesulfonates (BABS),phenylalkanesulfonates, alpha-olefinsulfonates (AOS), olefin sulfonates,alkene sulfonates, alkane-2,3-diylbis(sulfates), hydroxyalkanesulfonatesand disulfonates, alkyl sulfates (AS) such as sodium dodecyl sulfate(SDS), fatty alcohol sulfates (FAS), primary alcohol sulfates (PAS),alcohol ethersulfates (AES or AEOS or FES, also known as alcoholethoxysulfates or fatty alcohol ether sulfates), secondaryalkanesulfonates (SAS), paraffin sulfonates (PS), ester sulfonates,sulfonated fatty acid glycerol esters, alpha-sulfo fatty acid methylesters (alpha-SFMe or SES) including methyl ester sulfonate (MES),alkyl- or alkenylsuccinic acid, dodecenyl/tetradecenyl succinic acid(DTSA), fatty acid derivatives of amino acids, diesters and monoestersof sulfo-succinic acid or soap, and combinations thereof.

When included therein the detergent will usually comprise from about 0%to about 10% by weight of a cationic surfactant. Non-limiting examplesof cationic surfactants include alklydimethylethanolamine quat (ADMEAQ),cetyltrimethylammonium bromide (CTAB), dimethyldistearylammoniumchloride (DSDMAC), and alkylbenzyldimethylammonium, alkyl quaternaryammonium compounds, alkoxylated quaternary ammonium (AQA) compounds, andcombinations thereof.

When included therein the detergent will usually comprise from about0.2% to about 40% by weight of a non-ionic surfactant, for example fromabout 0.5% to about 30%, in particular from about 1% to about 20%, fromabout 3% to about 10%, such as from about 3% to about 5%, or from about8% to about 12%. Non-limiting examples of non-ionic surfactants includealcohol ethoxylates (AE or AEO), alcohol propoxylates, propoxylatedfatty alcohols (PFA), alkoxylated fatty acid alkyl esters, such asethoxylated and/or propoxylated fatty acid alkyl esters, alkylphenolethoxylates (APE), nonylphenol ethoxylates (NPE), alkylpolyglycosides(APG), alkoxylated amines, fatty acid monoethanolamides (FAM), fattyacid diethanolamides (FADA), ethoxylated fatty acid monoethanolamides(EFAM), propoxylated fatty acid monoethanolamides (PFAM), polyhydroxyalkyl fatty acid amides, or N-acyl N-alkyl derivatives of glucosamine(glucamides, GA, or fatty acid glucamide, FAGA), as well as productsavailable under the trade names SPAN and TWEEN, and combinationsthereof.

When included therein the detergent will usually comprise from about 0%to about 10% by weight of a semipolar surfactant. Non-limiting examplesof semipolar surfactants include amine oxides (AO) such asalkyldimethylamineoxide, N-(coco alkyl)-N,N-dimethylamine oxide andN-(tallow-alkyl)-N,N-bis(2-hydroxyethyl)amine oxide, fatty acidalkanolamides and ethoxylated fatty acid alkanolamides, and combinationsthereof.

When included therein the detergent will usually comprise from about 0%to about 10% by weight of a zwitterionic surfactant. Non-limitingexamples of zwitterionic surfactants include betaine,alkyldimethylbetaine, sulfobetaine, and combinations thereof.

Hydrotropes

A hydrotrope is a compound that solubilises hydrophobic compounds inaqueous solutions (or oppositely, polar substances in a non-polarenvironment). Typically, hydrotropes have both hydrophilic and ahydrophobic character (so-called amphiphilic properties as known fromsurfactants); however, the molecular structure of hydrotropes generallydo not favor spontaneous self-aggregation, see e.g. review by Hodgdonand Kaler (2007), Current Opinion in Colloid & Interface Science 12:121-128. Hydrotropes do not display a critical concentration above whichself-aggregation occurs as found for surfactants and lipids formingmiceller, lamellar or other well defined meso-phases. Instead, manyhydrotropes show a continuous-type aggregation process where the sizesof aggregates grow as concentration increases. However, many hydrotropesalter the phase behavior, stability, and colloidal properties of systemscontaining substances of polar and non-polar character, includingmixtures of water, oil, surfactants, and polymers. Hydrotropes areclassically used across industries from pharma, personal care, food, totechnical applications. Use of hydrotropes in detergent compositionsallow for example more concentrated formulations of surfactants (as inthe process of compacting liquid detergents by removing water) withoutinducing undesired phenomena such as phase separation or high viscosity.

The detergent may comprise from about 0 to about 5% by weight, such asfrom about 0.5 to about 5%, or about 3% to about 5%, of a hydrotrope.Any hydrotrope known in the art for use in detergents may be utilized.Non-limiting examples of hydrotropes include sodium benzene sulfonate,sodium p-toluene sulfonate (STS), sodium xylene sulfonate (SXS), sodiumcumene sulfonate (SCS), sodium cymene sulfonate, amine oxides, alcoholsand polyglycolethers, sodium hydroxynaphthoate, sodiumhydroxynaphthalene sulfonate, sodium ethylhexyl sulfate, andcombinations thereof.

Builders and Co-Builders

The detergent composition may comprise from about 0 to about 65% byweight, such as about 5% to about 45% of a detergent builder orco-builder, or a mixture thereof. In a dish wash detergent, the level ofbuilder is typically from about 40 to about 65%, particularly from about50 to about 65%. The builder and/or co-builder may particularly be achelating agent that forms water-soluble complexes with Ca and Mg. Anybuilder and/or co-builder known in the art for use in laundry detergentsmay be utilized. Non-limiting examples of builders include zeolites,diphosphates (pyrophosphates), triphosphates such as sodium triphosphate(STP or STPP), carbonates such as sodium carbonate, soluble silicatessuch as sodium metasilicate, layered silicates (e.g., SKS-6 fromHoechst), ethanolamines such as 2-aminoethan-1-ol (MEA), diethanolamine(DEA, also known as iminodiethanol), triethanolamine (TEA, also known as2,2′,2″-nitrilotriethanol), and carboxymethyl inulin (CMI), andcombinations thereof.

The detergent composition may also comprise from about 0 to about 20% byweight, such as about 5% to about 10%, of a detergent co-builder, or amixture thereof. The detergent composition may include a co-builderalone, or in combination with a builder, for example a zeolite builder.Non-limiting examples of co-builders include homopolymers ofpolyacrylates or copolymers thereof, such as poly(acrylic acid) (PAA) orcopoly(acrylic acid/maleic acid) (PAA/PMA). Further non-limitingexamples include citrate, chelators such as aminocarboxylates,aminopolycarboxylates and phosphonates, and alkyl- or alkenylsuccinicacid. Additional specific examples include 2,2′,2″-nitrilotriacetic acid(NTA), ethylenediaminetetraacetic acid (EDTA),diethylenetriaminepentaacetic acid (DTPA), iminodisuccinic acid (IDS),ethylenediamine-N,N′-disuccinic acid (EDDS), methylglycinediacetic acid(MGDA), glutamic acid-N,N-diacetic acid (GLDA),1-hydroxyethane-1,1-diphosphonic acid (HEDP),ethylenediaminetetra-(methylenephosphonic acid) (EDTMPA),diethylenetriaminepentakis(methylenephosphonic acid) (DTPMPA or DTMPA),N-(2-hydroxyethyl)iminodiacetic acid (EDG), aspartic acid-N-monoaceticacid (ASMA), aspartic acid-N,N-diacetic acid (ASDA), asparticacid-N-monopropionic acid (ASMP), iminodisuccinic acid (IDA),N-(2-sulfomethyl)-aspartic acid (SMAS), N-(2-sulfoethyl)-aspartic acid(SEAS), N-(2-sulfomethyl)-glutamic acid (SMGL),N-(2-sulfoethyl)-glutamic acid (SEGL), N-methyliminodiacetic acid(MIDA), α-alanine-N, N-diacetic acid (α-ALDA), serine-N, N-diacetic acid(SEDA), isoserine-N, N-diacetic acid (ISDA), phenylalanine-N, N-diaceticacid (PHDA), anthranilic acid-N, N-diacetic acid (ANDA), sulfanilicacid-N, N-diacetic acid (SLDA), taurine-N, N-diacetic acid (TUDA) andsulfomethyl-N, N-diacetic acid (SMDA),N-(2-hydroxyethyl)-ethylidenediamine-N, N′,N′-triacetate (HEDTA),diethanolglycine (DEG), diethylenetriamine penta(methylenephosphonicacid) (DTPMP), aminotris(methylenephosphonic acid) (ATMP), andcombinations and salts thereof. Further exemplary builders and/orco-builders are described in, e.g., WO 09/102854, U.S. Pat. No.5,977,053

Bleaching Systems

The detergent may comprise from about 0 to about 50% by weight, such asfrom about 0.1% to about 25%, of a bleaching system. Any bleachingsystem known in the art for use in laundry detergents may be utilized.Suitable bleaching system components include bleaching catalysts,photobleaches, bleach activators, sources of hydrogen peroxide such assodium percarbonate and sodium perborates, preformed peracids andmixtures thereof. Suitable preformed peracids include, but are notlimited to, peroxycarboxylic acids and salts, percarbonic acids andsalts, perimidic acids and salts, peroxymonosulfuric acids and salts,for example, Oxone (R), and mixtures thereof. Non-limiting examples ofbleaching systems include peroxide-based bleaching systems, which maycomprise, for example, an inorganic salt, including alkali metal saltssuch as sodium salts of perborate (usually mono- or tetra-hydrate),percarbonate, persulfate, perphosphate, persilicate salts, incombination with a peracid-forming bleach activator. The term bleachactivator is meant herein as a compound which reacts with peroxygenbleach like hydrogen peroxide to form a peracid. The peracid thus formedconstitutes the activated bleach. Suitable bleach activators to be usedherein include those belonging to the class of esters amides, imides oranhydrides. Suitable examples are tetracetylethylene diamine (TAED),sodium 4-[(3,5,5-trimethylhexanoyl)oxy]benzene sulfonate (ISONOBS),diperoxy dodecanoic acid, 4-(dodecanoyloxy)benzenesulfonate (LOBS),4-(decanoyloxy)benzenesulfonate, 4-(decanoyloxy)benzoate (DOBS),4-(nonanoyloxy)-benzenesulfonate (NOBS), and/or those disclosed inWO98/17767. A particular family of bleach activators of interest wasdisclosed in EP624154 and particulary preferred in that family is acetyltriethyl citrate (ATC). ATC or a short chain triglyceride like triacetinhas the advantage that it is environmental friendly as it eventuallydegrades into citric acid and alcohol. Furthermore acetyl triethylcitrate and triacetin has a good hydrolytical stability in the productupon storage and it is an efficient bleach activator. Finally ATCprovides a good building capacity to the laundry additive.Alternatively, the bleaching system may comprise peroxyacids of, forexample, the amide, imide, or sulfone type. The bleaching system mayalso comprise peracids such as 6-(phthalimido)peroxyhexanoic acid (PAP).The bleaching system may also include a bleach catalyst. In someembodiments the bleach component may be an organic catalyst selectedfrom the group of organic catalysts having the following formulae:

(iii) and mixtures thereof; wherein each R¹ is independently a branchedalkyl group containing from about 9 to about 24 carbons or linear alkylgroup containing from about 11 to about 24 carbons, preferably each R¹is independently a branched alkyl group containing from about 9 to about18 carbons or linear alkyl group containing from about 11 to about 18carbons, more preferably each R¹ is independently selected from thegroup of 2-propylheptyl, 2-butyloctyl, 2-pentylnonyl, 2-hexyldecyl,n-dodecyl, n-tetradecyl, n-hexadecyl, n-octadecyl, iso-nonyl, iso-decyl,iso-tridecyl and iso-pentadecyl. Other exemplary bleaching systems aredescribed, e.g. in WO2007/087258, WO2007/087244, WO2007/087259 andWO2007/087242. Suitable photobleaches may for example be sulfonated zincphthalocyanine

Polymers

The detergent may comprise from about 0 to about 10% by weight, such asfrom about 0.5 to about 5%, from about 2 to about 5%, from about 0.5 toabout 2% or from about 0.2 to about 1% of a polymer. Any polymer knownin the art for use in detergents may be utilized. The polymer mayfunction as a co-builder as mentioned above, or may provideantiredeposition, fiber protection, soil release, dye transferinhibition, grease cleaning and/or anti-foaming properties. Somepolymers may have more than one of the above-mentioned properties and/ormore than one of the below-mentioned motifs. Exemplary polymers include(carboxymethyl)cellulose (CMC), poly(vinyl alcohol) (PVA),poly(vinylpyrrolidone) (PVP), poly(ethyleneglycol) or poly(ethyleneoxide) (PEG), ethoxylated poly(ethyleneimine), carboxymethyl inulin(CMI), and polycarboxylates such as PAA, PAA/PMA, poly-aspartic acid,and lauryl methacrylate/acrylic acid copolymers, hydrophobicallymodified CMC (HM-CMC) and silicones, copolymers of terephthalic acid andoligomeric glycols, copolymers of poly(ethylene terephthalate) andpoly(oxyethene terephthalate) (PET-POET), PVP, poly(vinylimidazole)(PVI), poly(vinylpyridine-N-oxide) (PVPO or PVPNO) andpolyvinylpyrrolidone-vinylimidazole (PVPVI). Further exemplary polymersinclude sulfonated polycarboxylates, polyethylene oxide andpolypropylene oxide (PEO-PPO) and diquaternium ethoxy sulfate. Otherexemplary polymers are disclosed in, e.g., WO 2006/130575. Salts of theabove-mentioned polymers are also contemplated.

Fabric Hueing Agents

The detergent compositions of the present disclosure may also comprisefabric hueing agents such as dyes or pigments, which when formulated indetergent compositions can deposit onto a fabric when said fabric iscontacted with a wash liquor comprising said detergent compositions andthus altering the tint of said fabric through absorption/reflection ofvisible light. Fluorescent whitening agents emit at least some visiblelight. In contrast, fabric hueing agents alter the tint of a surface asthey absorb at least a portion of the visible light spectrum. Suitablefabric hueing agents include dyes and dye-clay conjugates, and may alsoinclude pigments. Suitable dyes include small molecule dyes andpolymeric dyes. Suitable small molecule dyes include small molecule dyesselected from the group of dyes falling into the Colour Index (C.I.)classifications of Direct Blue, Direct Red, Direct Violet, Acid Blue,Acid Red, Acid Violet, Basic Blue, Basic Violet and Basic Red, ormixtures thereof, for example as described in WO2005/03274,WO2005/03275, WO2005/03276 and EP1876226 (hereby incorporated byreference). The detergent composition preferably comprises from about0.00003 wt % to about 0.2 wt %, from about 0.00008 wt % to about 0.05 wt%, or even from about 0.0001 wt % to about 0.04 wt % fabric hueingagent. The composition may comprise from about 0.0001 wt % to about 0.2wt % fabric hueing agent, this may be especially preferred when thecomposition is in the form of a unit dose pouch. Suitable hueing agentsare also disclosed in, e.g. WO 2007/087257 and WO2007/087243.

Additional Enzymes

The detergent additive as well as the detergent composition may compriseone or more [additional] enzymes such as a xanthan lyase, protease,lipase, cutinase, an amylase, lichenase, carbohydrase, cellulase,pectinase, mannanase, arabinase, galactanase, xylanase, oxidase, e.g., alaccase, and/or peroxidase.

In general, the properties of the selected enzyme(s) should becompatible with the selected detergent, (i.e., pH-optimum, compatibilitywith other enzymatic and non-enzymatic ingredients, etc.), and theenzyme(s) should be present in effective amounts.

Cellulases:

Suitable cellulases include those of bacterial or fungal origin.Chemically modified or protein engineered mutants are included. Suitablecellulases include cellulases from the genera Bacillus, Pseudomonas,Humicola, Fusarium, Thielavia, Acremonium, e.g., the fungal cellulasesproduced from Humicola insolens, Myceliophthora thermophila and Fusariumoxysporum disclosed in U.S. Pat. Nos. 4,435,307, 5,648,263, 5,691,178,5,776,757 and WO 89/09259.

Especially suitable cellulases are the alkaline or neutral cellulaseshaving color care benefits. Examples of such cellulases are cellulasesdescribed in EP 0 495 257, EP 0 531 372, WO 96/11262, WO 96/29397, WO98/08940. Other examples are cellulase variants such as those describedin WO 94/07998, EP 0 531 315, U.S. Pat. Nos. 5,457,046, 5,686,593,5,763,254, WO 95/24471, WO 98/12307 and PCT/DK98/00299.

Example of cellulases exhibiting endo-beta-1,4-glucanase activity (EC3.2.1.4) are those having described in WO02/099091.

Other examples of cellulases include the family 45 cellulases describedin WO96/29397, and especially variants thereof having substitution,insertion and/or deletion at one or more of the positions correspondingto the following positions in SEQ ID NO: 8 of WO 02/099091: 2, 4, 7, 8,10, 13, 15, 19, 20, 21, 25, 26, 29, 32, 33, 34, 35, 37, 40, 42, 42a, 43,44, 48, 53, 54, 55, 58, 59, 63, 64, 65, 66, 67, 70, 72, 76, 79, 80, 82,84, 86, 88, 90, 91, 93, 95, 95d, 95h, 95j, 97, 100, 101, 102, 103, 113,114, 117, 119, 121, 133, 136, 137, 138, 139, 140a, 141, 143a, 145, 146,147, 150e, 150j, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160c,160e, 160k, 161, 162, 164, 165, 168, 170, 171, 172, 173, 175, 176, 178,181, 183, 184, 185, 186, 188, 191, 192, 195, 196, 200, and/or 20,preferably selected among P19A, G20K, Q44K, N48E, Q119H or Q146 R.

Commercially available cellulases include Celluzyme™, and Carezyme™(Novozymes A/S), Clazinase™, and Puradax HA™ (Genencor InternationalInc.), and KAC-500(B)™ (Kao Corporation).

Proteases:

The additional enzyme may be another protease or protease variant. Theprotease may be of animal, vegetable or microbial origin, includingchemically or genetically modified mutants. Microbial origin ispreferred. It may be an alkaline protease, such as a serine protease ora metalloprotease. A serine protease may for example be of the S1family, such as trypsin, or the S8 family such as subtilisin. Ametalloproteases protease may for example be a thermolysin from e.g.family M4, M5, M7 or M8.

The term “subtilases” refers to a sub-group of serine protease accordingto Siezen et al., Protein Engng. 4 (1991) 719-737 and Siezen et al.Protein Science 6 (1997) 501-523. Serine proteases are a subgroup ofproteases exemplified by having a serine in the active site, which formsa covalent adduct with the substrate. The subtilases may be divided into6 sub-divisions, i.e. the Subtilisin family, the Thermitase family, theProteinase K family, the Lantibiotic peptidase family, the Kexin familyand the Pyrolysin family. In one aspect of the disclosure the proteasemay be a subtilase, such as a subtilisin or a variant hereof. Furtherthe subtilases (and the serine proteases) are exemplified by having twoactive site amino acid residues apart from the serine, namely ahistidine and an aspartic acid residue.

Examples of subtilisins are those derived from Bacillus such assubtilisin lentus, Bacillus lentus, subtilisin Novo, subtilisinCarlsberg, Bacillus licheniformis, subtilisin BPN′, subtilisin 309,subtilisin 147 and subtilisin 168 described in WO 89/06279 and proteasePD138 (WO 93/18140). Additional serine protease examples are describedin WO 98/020115, WO 01/44452, WO 01/58275, WO 01/58276, WO 03/006602 andWO 04/099401. An example of a subtilase variants may be those havingmutations in any of the positions: 3, 4, 9, 15, 27, 36, 68, 76, 87, 95,96, 97, 98, 99, 100, 101, 102, 103, 104, 106, 118, 120, 123, 128, 129,130, 160, 167, 170, 194, 195, 199, 205, 217, 218, 222, 232, 235, 236,245, 248, 252 and 274 using the BPN′ numbering. More preferred thesubtilase variants may comprise the mutations: S3T, V4I, S9R, A15T,K27R, *36D, V68A, N76D, N87S,R, *97E, A98S, S99G,D,A, S99AD, S101G,M,RS103A, V104I,Y,N, S106A, G118V,R, H120D,N, N123S, S128L, P129Q, S130A,G160D, Y167A, R170S, A194P, G195E, V199M, V205I, L217D, N218D, M222S,A232V, K235L, Q236H, Q245R, N252K, T274A (using BPN′ numbering). Afurther preferred protease is the alkaline protease from Bacillus lentusDSM 5483, as described for example in WO 95/23221, and variants thereofwhich are described in WO 92/21760, WO 95/23221, EP 1921147 and EP1921148.

Examples of trypsin-like proteases are trypsin (e.g. of porcine orbovine origin) and the Fusarium protease described in WO 89/06270 and WO94/25583. Examples of useful proteases are the variants described in WO92/19729, WO 98/20115, WO 98/20116, and WO 98/34946, especially thevariants with substitutions in one or more of the following positions:27, 36, 57, 76, 87, 97, 101, 104, 120, 123, 167, 170, 194, 206, 218,222, 224, 235, and 274.

Examples of metalloproteases are the neutral metalloprotease asdescribed in WO 07/044993.

Preferred commercially available protease enzymes include Alcalase™,Coronase™, Duralase™, Durazym™, Esperase™, Everlase™, Kannase™,Liquanase™, Liquanase Ultra™, Ovozyme™, Polarzyme™, Primase™, Relase™,Savinase™ and Savinase Ultra™, (Novozymes A/S), Axapem™ (Gist-BrocasesN.V.), BLAP and BLAP X (Henkel AG & Co. KGaA), Excellase™, FN2™, FN3™,FN4™, Maxaca™, Maxapem™, Maxatase™, Properase™, Purafast™, Purafect™,Purafect OxP™, Purafect Prime™ and Puramax™ (Genencor int.).

Lipases and Cutinases:

Suitable lipases and cutinases include those of bacterial or fungalorigin. Chemically modified or protein engineered mutant enzymes areincluded. Examples include lipase from Thermomyces, e.g. from T.lanuginosus (previously named Humicola lanuginosa) as described inEP258068 and EP305216, cutinase from Humicola, e.g. H. insolens(WO96/13580), lipase from strains of Pseudomonas (some of these nowrenamed to Burkholderia), e.g. P. alcaligenes or P. pseudoalcaligenes(EP218272), P. cepacia (EP331376), P. sp. strain SD705 (WO95/06720 &WO96/27002), P. wisconsinensis (WO96/12012), GDSL-type Streptomyceslipases (WO10/065455), cutinase from Magnaporthe grisea (WO10/107560),cutinase from Pseudomonas mendocina (U.S. Pat. No. 5,389,536), lipasefrom Thermobifida fusca (WO11/084412), Geobacillus stearothermophiluslipase (WO11/084417), lipase from Bacillus subtilis (WO11/084599), andlipase from Streptomyces griseus (WO11/150157) and S. pristinaespiralis(WO12/137147).

Further examples are lipases sometimes referred to as acyltransferasesor perhydrolases, e.g. acyltransferases with homology to Candidaantarctica lipase A (WO10/111143), acyltransferase from Mycobacteriumsmegmatis (WO05/56782), perhydrolases from the CE 7 family (WO09/67279),and variants of the M. smegmatis perhydrolase in particular the S54Vvariant used in the commercial product Gentle Power Bleach from HuntsmanTextile Effects Pte Ltd (WO10/100028).

Other examples are lipase variants such as those described in EP407225,WO92/05249, WO94/01541, WO94/25578, WO95/14783, WO95/30744, WO95/35381,WO95/22615, WO96/00292, WO97/04079, WO97/07202, WO00/34450, WO00/60063,WO01/92502, WO07/87508 and WO09/109500.

Preferred commercial lipase products include Lipolase™, Lipex™; Lipolex™and Lipoclean™ (Novozymes A/S), Lumafast (originally from Genencor) andLipomax (originally from Gist-Brocades).

Amylases

The amylase may be an alpha-amylase, a beta-amylase or a glucoamylaseand may be of bacterial or fungal origin. Chemically modified or proteinengineered mutants are included. Amylases include, for example,alpha-amylases obtained from Bacillus, e.g., a special strain ofBacillus licheniformis, described in more detail in GB 1,296,839.

Examples of amylases are those having SEQ ID NO: 3 in WO 95/10603 orvariants having about 90% sequence identity to SEQ ID NO: 3 thereof.Preferred variants are described in WO 94/02597, WO 94/18314, WO97/43424 and SEQ ID NO: 4 of WO 99/019467, such as variants withsubstitutions in one or more of the following positions: 15, 23, 105,106, 124, 128, 133, 154, 156, 178, 179, 181, 188, 190, 197, 201, 202,207, 208, 209, 211, 243, 264, 304, 305, 391, 408, and 444 of SEQ ID NO:3 in WO 95/10603.

Further amylases which can be used are amylases having SEQ ID NO: 6 inWO 02/010355 or variants thereof having about 90% sequence identity toSEQ ID NO: 6. Preferred variants of SEQ ID NO: 6 are those having adeletion in positions 181 and 182 and a substitution in position 193.

Other amylase examples are hybrid alpha-amylase comprising residues 1-33of the alpha-amylase derived from B. amyloliquefaciens shown in SEQ IDNO: 6 of WO 2006/066594 and residues 36-483 of the B. licheniformisalpha-amylase shown in SEQ ID NO: 4 of WO 2006/066594 or variants havingabout 90% sequence identity thereof. Preferred variants of this hybridalpha-amylase are those having a substitution, a deletion or aninsertion in one of more of the following positions: G48, T49, G107,H156, A181, N190, M197, I201, A209 and Q264. Most preferred variants ofthe hybrid alpha-amylase comprising residues 1-33 of the alpha-amylasederived from B. amyloliquefaciens shown in SEQ ID NO: 6 of WO2006/066594 and residues 36-483 of SEQ ID NO: 4 are those having thesubstitutions: M197T; H156Y+A181T+N190F+A209V+Q264S; orG48+T49+G107+H156+A181+N190+I201+A209+Q264.

Further amylase examples are amylases having SEQ ID NO: 6 in WO99/019467 or variants thereof having about 90% sequence identity to SEQID NO: 6. Preferred variants of SEQ ID NO: 6 are those having asubstitution, a deletion or an insertion in one or more of the followingpositions: R181, G182, H183, G184, N195, I206, E212, E216 and K269.Particularly preferred amylases are those having deletion in positionsG182 and H183 or positions H183 and G184.

Additional amylases are those having SEQ ID NO: 1, SEQ ID NO: 2 or SEQID NO: 7 of WO 96/023873 or variants thereof having about 90% sequenceidentity to SEQ ID NO: 1, SEQ ID NO: 2 or SEQ ID NO: 7. Preferredvariants of SEQ ID NO: 1, SEQ ID NO: 2 or SEQ ID NO: 7 are those havinga substitution, a deletion or an insertion in one or more of thefollowing positions: 140, 181, 182, 183, 184, 195, 206, 212, 243, 260,269, 304 and 476. More preferred variants are those having a deletion inpositions 182 and 183 or positions 183 and 184. Most preferred amylasevariants of SEQ ID NO: 1, SEQ ID NO: 2 or SEQ ID NO: 7 are those havinga deletion in positions 183 and 184 and a substitution in positions 140,195, 206, 243, 260, 304 and 476.

Other amylases which can be used are amylases having SEQ ID NO: 2 of WO08/153815, SEQ ID NO: 10 in WO 01/66712 or variants thereof having about90% sequence identity to SEQ ID NO: 2 of WO 08/153815 or about 90%sequence identity to SEQ ID NO: 10 in WO 01/66712. Preferred variants ofSEQ ID NO: 10 in WO 01/66712 are those having a substitution, a deletionor an insertion in one of more of the following positions: 176, 177,178, 179, 190, 201, 207, 211 and 264.

Further amylases which can be used are amylases having SEQ ID NO: 2 ofWO 09/061380 or variants thereof having about 90% sequence identity toSEQ ID NO: 2. Preferred variants of SEQ ID NO: 2 are those having asubstitution, a deletion or an insertion in one of more of the followingpositions: Q87, Q98, S125, N128, T131, T165, K178, R180, 5181, T182,G183, M201, F202, N225, 5243, N272, N282, Y305, R309, D319, Q320, Q359,K444 and G475. More preferred variants of SEQ ID NO: 2 are those havingthe substitution in one of more of the following positions: Q87E,R,Q98R, S125A, N128C, T131I, T165I, K178L, T182G, M201L, F202Y, N225E,R,N272E,R, S243Q,A,E,D, Y305R, R309A, Q320R, Q359E, K444E and G475K and/ordeletion in position R180 and/or S181. Most preferred amylase variantsof SEQ ID NO: 2 are those having the substitutions:

N128C+K178L+T182G+Y305R+G475K;

N128C+K178L+T182G+F202Y+Y305R+D319T+G475K;

S125A+N128C+K178L+T182G+Y305R+G475K; or

S125A+N128C+T131I+T165I+K178L+T182G+Y305R+G475K wherein the variantoptionally further comprises a substitution at position 243 and/or adeletion at position 180 and/or position 181.

Other amylases are variants of SEQ ID NO: 1 of WO 2016/203064 having atleast 75% sequence identity to SEQ ID NO: 1 thereof. Preferred variantsare variants comprising a modification in one or more positionscorresponding to positions 1, 54, 56, 72, 109, 113, 116, 134, 140, 159,167, 169, 172, 173, 174, 181, 182, 183, 184, 189, 194, 195, 206, 255,260, 262, 265, 284, 289, 304, 305, 347, 391, 395, 439, 469, 444, 473,476, or 477 of SEQ ID NO: 1, wherein said alpha-amylase variant has asequence identity of at least 75% but less than 100% to SEQ ID NO: 1.

Other examples of amylases are the alpha-amylase having SEQ ID NO: 12 inWO01/66712 or a variant having at least about 90%, such as at leastabout 95%, sequence identity to SEQ ID NO: 12. Preferred amylasevariants are those having a substitution, a deletion or an insertion inone of more of the following positions of SEQ ID NO: 12 in WO01/66712:R28, R118, N174; R181, G182, D183, G184, G186, W189, N195, M202, Y298,N299, K302, 5303, N306, R310, N314; R320, H324, E345, Y396, R400, W439,R444, N445, K446, Q449, R458, N471, N484. Particular preferred amylasesinclude variants having a deletion of D183 and G184 and having thesubstitutions R118K, N195F, R320K and R458K, and a variant additionallyhaving substitutions in one or more position selected from the group:M9, G149, G182, G186, M202, T257, Y295, N299, M323, E345 and A339, mostpreferred a variant that additionally has substitutions in all thesepositions.

Commercially available amylases are Duramyl™, Termamyl™, Fungamyl™Stainzyme™, Stainzyme Plus™, Natalase™ and BAN™ (Novozymes A/S),Rapidase™ and Purastar™ (from Genencor International Inc.).

Peroxidases/Oxidases: Suitable peroxidases/oxidases include those ofplant, bacterial or fungal origin. Chemically modified or proteinengineered mutants are included. Examples of useful peroxidases includeperoxidases from Coprinus, e.g., from C. cinereus, and variants thereofas those described in WO 93/24618, WO 95/10602, and WO 98/15257.Commercially available peroxidases include Guardzyme™ (Novozymes A/S).

The detergent enzyme(s) may be included in a detergent composition byadding separate additives containing one or more enzymes, or by adding acombined additive comprising all of these enzymes. A detergent additiveof the disclosure, i.e., a separate additive or a combined additive, canbe formulated, for example, as a granulate, liquid, slurry, etc.Preferred detergent additive formulations are granulates, in particularnon-dusting granulates, liquids, in particular stabilized liquids, orslurries.

Non-dusting granulates may be produced, e.g., as disclosed in U.S. Pat.Nos. 4,106,991 and 4,661,452 and may optionally be coated by methodsknown in the art. Examples of waxy coating materials are poly(ethyleneoxide) products (polyethyleneglycol, PEG) with mean molar weights offrom about 1000 to about 20000; ethoxylated nonylphenols having fromabout 16 to about 50 ethylene oxide units; ethoxylated fatty alcohols inwhich the alcohol contains from about 12 to about 20 carbon atoms and inwhich there are from about 15 to about 80 ethylene oxide units; fattyalcohols; fatty acids; and mono- and di- and triglycerides of fattyacids. Examples of film-forming coating materials suitable forapplication by fluid bed techniques are given in GB 1483591. Liquidenzyme preparations may, for instance, be stabilized by adding a polyolsuch as propylene glycol, a sugar or sugar alcohol, lactic acid or boricacid according to established methods. Protected enzymes may be preparedaccording to the method disclosed in EP 238,216.

Adjunct Materials

Any detergent components known in the art for use in laundry detergentsmay also be utilized. Other optional detergent components includeanti-corrosion agents, anti-shrink agents, anti-soil redepositionagents, anti-wrinkling agents, bactericides, binders, corrosioninhibitors, disintegrants/disintegration agents, dyes, enzymestabilizers (including boric acid, borates, CMC, and/or polyols such aspropylene glycol), fabric conditioners including clays,fillers/processing aids, fluorescent whitening agents/opticalbrighteners, foam boosters, foam (suds) regulators, perfumes,soil-suspending agents, softeners, suds suppressors, tarnish inhibitors,and wicking agents, either alone or in combination. Any ingredient knownin the art for use in laundry detergents may be utilized. The choice ofsuch ingredients is well within the skill of the artisan.

Dispersants:

The detergent compositions of the present disclosure can also containdispersants. In particular, powdered detergents may comprisedispersants. Suitable water-soluble organic materials include the homo-or co-polymeric acids or their salts, in which the polycarboxylic acidcomprises at least two carboxyl radicals separated from each other bynot more than two carbon atoms. Suitable dispersants are for exampledescribed in Powdered Detergents, Surfactant science series volume 71,Marcel Dekker, Inc.

Dye Transfer Inhibiting Agents:

The detergent compositions of the present disclosure may also includeone or more dye transfer inhibiting agents. Suitable polymeric dyetransfer inhibiting agents include, but are not limited to,polyvinylpyrrolidone polymers, polyamine N-oxide polymers, copolymers ofN-vinylpyrrolidone and N-vinylimidazole, polyvinyloxazolidones andpolyvinylimidazoles or mixtures thereof. When present in a subjectcomposition, the dye transfer inhibiting agents may be present at levelsfrom about 0.0001% to about 10%, from about 0.01% to about 5% or evenfrom about 0.1% to about 3% by weight of the composition.

Fluorescent Whitening Agent:

The detergent compositions of the present disclosure will preferablyalso contain additional components that may tint articles being cleaned,such as fluorescent whitening agent or optical brighteners. Wherepresent the brightener is preferably at a level of from about 0.01% toabout 0.5%. Any fluorescent whitening agent suitable for use in alaundry detergent composition may be used in the composition of thepresent disclosure. The most commonly used fluorescent whitening agentsare those belonging to the classes of diaminostilbene-sulphonic acidderivatives, diarylpyrazoline derivatives and bisphenyl-distyrylderivatives. Examples of the diaminostilbene-sulphonic acid derivativetype of fluorescent whitening agents include the sodium salts of:4,4′-bis-(2-diethanolamino-4-anilino-s-triazin-6-ylamino)stilbene-2,2′-disulphonate; 4,4′-bis-(2,4-dianilino-s-triazin-6-ylamino)stilbene-2,2′-disulphonate;4,4′-bis-(2-anilino-4(N-methyl-N-2-hydroxy-ethylamino)-s-triazin-6-ylamino)stilbene-2,2′-disulphonate,4,4′-bis-(4-phenyl-2,1,3-triazol-2-yl)stilbene-2,2′-disulphonate;4,4′-bis-(2-anilino-4(1-methyl-2-hydroxy-ethylamino)-s-triazin-6-ylamino)stilbene-2,2′-disulphonate and2-(stilbyl-4″-naptho-1,2′:4,5)-1,2,3-trizole-2″-sulphonate. Preferredfluorescent whitening agents are Tinopal DMS and Tinopal CBS availablefrom Ciba-Geigy AG, Basel, Switzerland. Tinopal DMS is the disodium saltof 4,4′-bis-(2-morpholino-4 anilino-s-triazin-6-ylamino) stilbenedisulphonate. Tinopal CBS is the disodium salt of2,2′-bis-(phenyl-styryl) disulphonate. Also preferred are fluorescentwhitening agents is the commercially available Parawhite KX, supplied byParamount Minerals and Chemicals, Mumbai, India. Other fluorescerssuitable for use in the disclosure include the 1-3-diaryl pyrazolinesand the 7-alkylaminocoumarins. Suitable fluorescent brightener levelsinclude lower levels of from about 0.01, from about 0.05, from about 0.1or even from about 0.2 wt % to upper levels of about 0.5 or even about0.75 wt %.

Soil Release Polymers:

The detergent compositions of the present disclosure may also includeone or more soil release polymers which aid the removal of soils fromfabrics such as cotton and polyester based fabrics, in particular theremoval of hydrophobic soils from polyester based fabrics. The soilrelease polymers may for example be nonionic or anionic terephthaltebased polymers, polyvinyl caprolactam and related copolymers, vinylgraft copolymers, polyester polyamides see for example Chapter 7 inPowdered Detergents, Surfactant science series volume 71, Marcel Dekker,Inc. Another type of soil release polymers is amphiphilic alkoxylatedgrease cleaning polymers comprising a core structure and a plurality ofalkoxylate groups attached to that core structure. The core structuremay comprise a polyalkylenimine structure or a polyalkanolaminestructure as described in detail in WO 2009/087523 (hereby incorporatedby reference). Furthermore random graft co-polymers are suitable soilrelease polymers Suitable graft co-polymers are described in more detailin WO 2007/138054, WO 2006/108856 and WO 2006/113314 (herebyincorporated by reference). Other soil release polymers are substitutedpolysaccharide structures especially substituted cellulosic structuressuch as modified cellulose deriviatives such as those described in EP1867808 or WO 2003/040279 (both are hereby incorporated by reference).Suitable cellulosic polymers include cellulose, cellulose ethers,cellulose esters, cellulose amides and mixtures thereof. Suitablecellulosic polymers include anionically modified cellulose, nonionicallymodified cellulose, cationically modified cellulose, zwitterionicallymodified cellulose, and mixtures thereof. Suitable cellulosic polymersinclude methyl cellulose, carboxy methyl cellulose, ethyl cellulose,hydroxyl ethyl cellulose, hydroxyl propyl methyl cellulose, estercarboxy methyl cellulose, and mixtures thereof.

Anti-Redeposition Agents:

The detergent compositions of the present disclosure may also includeone or more anti-redeposition agents such as carboxymethylcellulose(CMC), polyvinyl alcohol (PVA), polyvinylpyrrolidone (PVP),polyoxyethylene and/or polyethyleneglycol (PEG), homopolymers of acrylicacid, copolymers of acrylic acid and maleic acid, and ethoxylatedpolyethyleneimines. The cellulose based polymers described under soilrelease polymers above may also function as anti-redeposition agents.

Other suitable adjunct materials include, but are not limited to,anti-shrink agents, anti-wrinkling agents, bactericides, binders,carriers, dyes, enzyme stabilizers, fabric softeners, fillers, foamregulators, hydrotropes, perfumes, pigments, sod suppressors, solvents,and structurants for liquid detergents and/or structure elasticizingagents.

Formulation of Detergent Products

The detergent composition may be in any convenient form, e.g., a bar, ahomogenous tablet, a tablet having two or more layers, a pouch havingone or more compartments, a regular or compact powder, a granule, apaste, a gel, or a regular, compact or concentrated liquid. There are anumber of detergent formulation forms such as layers (same or differentphases), pouches, as well as forms for machine dosing unit.

Pouches can be configured as single or multicompartments. It can be ofany form, shape and material which is suitable for hold the composition,e.g. without allowing the release of the composition from the pouchprior to water contact. The pouch is made from water soluble film whichencloses an inner volume. Said inner volume can be divided intocompartments of the pouch. Preferred films are polymeric materialspreferably polymers which are formed into a film or sheet. Preferredpolymers, copolymers or derivates thereof are selected polyacrylates,and water soluble acrylate copolymers, methyl cellulose, carboxy methylcellulose, sodium dextrin, ethyl cellulose, hydroxyethyl cellulose,hydroxypropyl methyl cellulose, malto dextrin, poly methacrylates, mostpreferably polyvinyl alcohol copolymers and, hydroxyprpyl methylcellulose (HPMC). Preferably the level of polymer in the film forexample PVA is at least about 60%. Preferred average molecular weightwill typically be from about 20,000 to about 150,000. Films can also beof blend compositions comprising hydrolytically degradable and watersoluble polymer blends such as polyactide and polyvinyl alcohol (knownunder the Trade reference M8630 as sold by Chris Craft In. Prod. OfGary, Ind., US) plus plasticisers like glycerol, ethylene glycerol,Propylene glycol, sorbitol and mixtures thereof. The pouches cancomprise a solid laundry cleaning composition or part components and/ora liquid cleaning composition or part components separated by the watersoluble film. The compartment for liquid components can be different incomposition than compartments containing solids. Ref: (US2009/0011970A1).

Detergent ingredients can be separated physically from each other bycompartments in water dissolvable pouches or in different layers oftablets. Thereby negative storage interaction between components can beavoided. Different dissolution profiles of each of the compartments canalso give rise to delayed dissolution of selected components in the washsolution.

A liquid or gel detergent, which is not unit dosed, may be aqueous,typically containing at least about 20% by weight and up to about 95%water, such as up to about 70% water, up to about 65% water, up to about55% water, up to about 45% water, up to about 35% water. Other types ofliquids, including without limitation, alkanols, amines, diols, ethersand polyols may be included in an aqueous liquid or gel. An aqueousliquid or gel detergent may contain from about 0 to about 30% organicsolvent. A liquid or gel detergent may be non-aqueous.

Laundry Soap Bars

The enzymes of the disclosure may be added to laundry soap bars and usedfor hand washing laundry, fabrics and/or textiles. The term laundry soapbar includes laundry bars, soap bars, combo bars, syndet bars anddetergent bars. The types of bar usually differ in the type ofsurfactant they contain, and the term laundry soap bar includes thosecontaining soaps from fatty acids and/or synthetic soaps. The laundrysoap bar has a physical form which is solid and not a liquid, gel or apowder at room temperature. The term solid is defined as a physical formwhich does not significantly change over time, i.e. if a solid object(e.g. laundry soap bar) is placed inside a container, the solid objectdoes not change to fill the container it is placed in. The bar is asolid typically in bar form but can be in other solid shapes such asround or oval.

The laundry soap bar may contain one or more additional enzymes,protease inhibitors such as peptide aldehydes (or hydrosulfite adduct orhemiacetal adduct), boric acid, borate, borax and/or phenylboronic acidderivatives such as 4-formylphenylboronic acid, one or more soaps orsynthetic surfactants, polyols such as glycerine, pH controllingcompounds such as fatty acids, citric acid, acetic acid and/or formicacid, and/or a salt of a monovalent cation and an organic anion whereinthe monovalent cation may be for example Na⁺, K⁺ or NH₄ ⁺ and theorganic anion may be for example formate, acetate, citrate or lactatesuch that the salt of a monovalent cation and an organic anion may be,for example, sodium formate.

The laundry soap bar may also contain complexing agents like EDTA andHEDP, perfumes and/or different type of fillers, surfactants e.g.anionic synthetic surfactants, builders, polymeric soil release agents,detergent chelators, stabilizing agents, fillers, dyes, colorants, dyetransfer inhibitors, alkoxylated polycarbonates, suds suppressers,structurants, binders, leaching agents, bleaching activators, clay soilremoval agents, anti-redeposition agents, polymeric dispersing agents,brighteners, fabric softeners, perfumes and/or other compounds known inthe art.

The laundry soap bar may be processed in conventional laundry soap barmaking equipment such as but not limited to: mixers, plodders, e.g a twostage vacuum plodder, extruders, cutters, logo-stampers, cooling tunnelsand wrappers. The disclosure is not limited to preparing the laundrysoap bars by any single method. The premix of the disclosure may beadded to the soap at different stages of the process. For example, thepremix containing a soap, an enzyme, optionally one or more additionalenzymes, a protease inhibitor, and a salt of a monovalent cation and anorganic anion may be prepared and the mixture is then plodded. Theenzyme and optional additional enzymes may be added at the same time asthe protease inhibitor for example in liquid form. Besides the mixingstep and the plodding step, the process may further comprise the stepsof milling, extruding, cutting, stamping, cooling and/or wrapping.

Method of Producing the Composition

The present disclosure also relates to methods of producing thecomposition. The method may be relevant for the (storage) stability ofthe detergent composition: e.g. Soap bar premix method WO2009155557.

Uses

The present disclosure is also directed to methods for using thedetergent compositions thereof. The present disclosure may be used forexample in any detergent application which requires the degradation ofxanthan gum.

Use to Degrade Xanthan Gum

Xanthan gum has been used as an ingredient in many consumer productsincluding foods and cosmetics and has found use in the oil industry.Therefore, the degradation of xanthan gum can result in improvedcleaning processes, such as the easier removal of stains containinggums, such as xanthan gum. Thus, the present disclosure is directed tothe use of detergent compositions comprising GH9 endoglucanases (e.g.variants described herein) of the disclosure to degrade xanthan gum. Thepresent disclosure is also directed to the use of xanthan lyases in thecompositions of the disclosure to degrade xanthan gum. An embodiment isthe use of a detergent composition comprising GH9 endoglucanases asdescribed herein (e.g. variants) together with xanthan lyases to degradexanthan gum. Degradation of xanthan gum can preferably be measured usingthe viscosity reduction assay (e.g., ViPr assay) or alternatively asdescribed in example 4 herein.

GH9 endoglucanase activity may alternatively be measured by assessmentof reducing ends on xanthan gum pre-treated with xanthan lyase using thecolorimetric assay developed by Lever (1972), Anal. Biochem. 47:273-279, 1972. A preferred embodiment is the use of about 0.1% xanthangum pre-treated with xanthan lyase. Degradation of xanthan gumpre-treated with xanthan lyase may be determined by calculatingdifference between blank and sample, wherein a difference of more thanabout 0.5 mAU, preferably more than about 0.6 mAU, more preferably morethan about 0.7 mAU or even more preferably more than about 0.8 mAU,shows degradation of xanthan gum pre-treated with xanthan lyase.

Xanthan lyase activity may alternatively be measured by assessment ofreducing ends liberated from xanthan gum using the colorimetric assaydeveloped by Lever (1972), Anal. Biochem. 47: 273-279, 1972. A preferredembodiment is the use of about 0.1% xanthan gum. Degradation of xanthangum may be determined by calculating difference between blank and samplewherein a difference of more than about 0.1 mAU, preferably more thanabout 0.15 mAU, more preferably more than about 0.2 mAU or even morepreferably more than about 0.25 mAU shows degradation of xanthan gum.

GH9 endoglucanase (e.g. variants of the present disclosure) and xanthanlyase activity may alternatively be measured by assessment of reducingends liberated from xanthan gum using the colorimetric assay developedby Lever (1972), Anal. Biochem. 47: 273-279, 1972. A preferredembodiment is the use of about 0.1% xanthan gum. Degradation of xanthangum may be determined by calculating difference between blank and samplewherein a difference of more than about 0.4 mAU, preferably more than0.5 mAU, more preferably more than 0.6 mAU or even more preferably morethan about 0.8 mAU shows degradation of xanthan gum.

The disclosure also relates to methods for degrading xanthan gumcomprising applying a detergent composition comprising one or more GH9endoglucanases described herein (e.g. variants) to xanthan gum. Anembodiment is a method for degrading xanthan gum comprising applying adetergent composition comprising one or more GH9 endoglucanases asdescribed herein (e.g. variants) together with one or more xanthanlyases to xanthan gum.

Use in Detergents

The present disclosure inter alia relates to the use of detergentcompositions comprising GH9 endoglucanases as described herein (e.g.variants) in cleaning processes such as the laundering of textiles andfabrics (e.g. household laundry washing and industrial laundry washing),as well as household and industrial hard surface cleaning, such as dishwash. For this, the GH9 endoglucanases (e.g. variants) may be added to adetergent composition comprising of one or more detergent components.

In some aspects GH9 endoglucanases as described herein (e.g. variants)may be used together with a xanthan lyase(s) in detergent compositionsfor cleaning processes such as the laundering of textiles and fabrics(e.g. household laundry washing and industrial laundry washing), as wellas household and industrial hard surface cleaning, such as dish wash.For this, the GH9 endoglucanases described herein (e.g. variants)together with a xanthan lyase(s) may be added to a detergent compositioncomprising of one or more detergent components.

The polypeptides described herein (e.g. variants) may be added to andthus become a component of a detergent composition. The detergentcomposition may be formulated, for example, as a hand or machine laundrydetergent composition for both household and industrial laundrycleaning, including a laundry additive composition suitable forpre-treatment of stained fabrics and a rinse added fabric softenercomposition, or be formulated as a detergent composition for use ingeneral household or industrial hard surface cleaning operations, or beformulated for hand or machine (both household and industrial)dishwashing operations. In a specific aspect, the present disclosurerelates to a detergent additive comprising a polypeptide as describedherein.

The disclosure also relates to methods for degrading xanthan gum on thesurface of a textile or hard surface, such as dish wash, comprisingapplying a detergent composition comprising one or more GH9endoglucanases as described herein (e.g. variants) to xanthan gum. Insome aspects, the disclosure relates a method for degrading xanthan gumon the surface of a textile or hard surface, such as dish wash,comprising applying a detergent composition comprising one or more GH9endoglucanases described herein (e.g. variants) together with one ormore xanthan lyases to xanthan gum. In some aspects, the disclosurerelates to a detergent composition comprising one or more detergentcomponents as described herein. Also encompassed is the use of GH9endoglucanases (e.g. variants) having an enzyme detergency benefit insaid detergent compositions.

It has been contemplated that the use of a GH9 endoglucanase asdescribed herein (e.g., a variant as described herein) alone gives anenzyme detergency benefit, preferably an enzyme detergency benefit onxanthan gum.

In some aspects, the disclosure relates to the use of a detergentcomposition comprising one or more detergent components and an isolatedGH9 endoglucanase described herein (e.g. a variant) together with axanthan lyase. In some aspects, the disclosure relates to the use of adetergent composition comprising one or more detergent components and anisolated GH9 endoglucanase (e.g. a variant) together with a xanthanlyase.

The Disclosure is Further Defined in the Following Paragraphs:

1. A detergent composition comprising an endoglucanase variant,comprising an alteration (e.g., a substitution, deletion or insertion)at one or more positions in a region selected from the group of:

i) region 1 corresponding to amino acids 95 to 105 of SEQ ID NO: 2,e.g., said alteration at one or more positions selected from the groupof positions: 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, whereinsaid positions correspond to amino acid positions of SEQ ID NO: 2 (e.g.,using the numbering of SEQ ID NO: 2),

ii) region 2 corresponding to amino acids 115 to 138 of SEQ ID NO: 2,e.g., said alteration at one or more positions selected from the groupof positions: 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125,126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, whereinsaid positions correspond to amino acid positions of SEQ ID NO: 2 (e.g.,using the numbering of SEQ ID NO: 2),

iii) region 3 corresponding to amino acids 210 to 251 of SEQ ID NO: 2,e.g., said alteration at one or more positions selected from the groupof positions: 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220,221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234,235, 236, 237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248,249, 250, 251, wherein said positions correspond to amino acid positionsof SEQ ID NO: 2 (e.g., using the numbering of SEQ ID NO: 2),

iv) region 4 corresponding to amino acids 267 to 301 of SEQ ID NO: 2,e.g., said alteration at one or more positions selected from the groupof positions: 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277,278, 279, 280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291,292, 293, 294, 295, 296, 297, 298, 299, 300, 301, wherein said positionscorrespond to amino acid positions of SEQ ID NO: 2 (e.g., using thenumbering of SEQ ID NO: 2),

v) region 5 corresponding to amino acids 339 to 361 of SEQ ID NO: 2,e.g., said alteration at one or more positions selected from the groupof positions: 339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349,350, 351, 352, 353, 354, 355, 356, 357, 358, 359, 360, 361, wherein saidpositions correspond to amino acid positions of SEQ ID NO: 2 (e.g.,using the numbering of SEQ ID NO: 2),

vi) region 6 corresponding to amino acids 547 to 595 of SEQ ID NO: 2,e.g., said alteration at one or more positions selected from the groupof positions: 547, 548, 549, 550, 551, 552, 553, 554, 555, 556, 557,558, 559, 560, 561, 562, 563, 564, 565, 566, 567, 568, 569, 570, 571,572, 573, 574, 575, 576, 577, 578, 579, 580, 581, 582, 583, 584, 585,586, 587, 588, 589, 590, 591, 592, 593, 594, 595, wherein said positionscorrespond to amino acid positions of SEQ ID NO: 2 (e.g., using thenumbering of SEQ ID NO: 2),

vii) region 7 corresponding to amino acids 612 to 660 of SEQ ID NO: 2,e.g., said alteration at one or more positions selected from the groupof positions: 612, 613, 614, 615, 616, 617, 618, 619, 620, 621, 622,623, 624, 625, 626, 627, 628, 629, 630, 631, 632, 633, 634, 635, 636,637, 638, 639, 640, 641, 642, 643, 644, 645, 646, 647, 648, 649, 650,651, 652, 653, 654, 655, 656, 657, 658, 659, 660, wherein said positionscorrespond to amino acid positions of SEQ ID NO: 2 (e.g., using thenumbering of SEQ ID NO: 2),

viii) region 8 corresponding to amino acids 806 to 828 of SEQ ID NO: 2,e.g., said alteration at one or more positions selected from the groupof positions: 806, 807, 808, 809, 810, 811, 812, 813, 814, 815, 816,817, 818, 819, 820, 821, 822, 823, 824, 825, 826, 827, 828, wherein saidpositions correspond to amino acid positions of SEQ ID NO: 2 (e.g.,using the numbering of SEQ ID NO: 2), and

ix) region 9 corresponding to amino acids 839 to 1042 of SEQ ID NO: 2,e.g., said alteration at one or more positions selected from the groupof positions: 839, 840, 841, 842, 843, 844, 845, 846, 847, 848, 849,850, 851, 852, 853, 854, 855, 856, 857, 858, 859, 860, 861, 862, 863,864, 865, 866, 867, 868, 869, 870, 871, 872, 873, 874, 875, 876, 877,878, 879, 880, 881, 882, 883, 884, 885, 886, 887, 888, 889, 890, 891,892, 893, 894, 895, 896, 897, 898, 899, 900, 901, 902, 903, 904, 905,906, 907, 908, 909, 910, 911, 912, 913, 914, 915, 916, 917, 918, 919,920, 921, 922, 923, 924, 925, 926, 927, 928, 929, 930, 931, 932, 933,934, 935, 936, 937, 938, 939, 940, 941, 942, 943, 944, 945, 946, 947,948, 949, 950, 951, 952, 953, 954, 955, 956, 957, 958, 959, 960, 961,962, 963, 964, 965, 966, 967, 968, 969, 970, 971, 972, 973, 974, 975,976, 977, 978, 979, 980, 981, 982, 983, 984, 985, 986, 987, 988, 989,990, 991, 992, 993, 994, 995, 996, 997, 998, 999, 1000, 1001, 1002,1003, 1004, 1005, 1006, 1007, 1008, 1009, 1010, 1011, 1012, 1013, 1014,1015, 1016, 1017, 1018, 1019, 1020, 1021, 1022, 1023, 1024, 1025, 1026,1027, 1028, 1029, 1030, 1031, 1032, 1033, 1034, 1035, 1036, 1037, 1038,1039, 1040, 1041, 1042, wherein said positions correspond to amino acidpositions of SEQ ID NO: 2 (e.g., using the numbering of SEQ ID NO: 2),

wherein said variant has at least about 60%, e.g., at least about 65%,at least about 70%, at least about 75%, at least about 80%, at leastabout 85%, at least about 90%, at least about 95%, at least about 96%,at least about 97%, at least about 98%, or at least about 99%, and lessthan about 100% sequence identity to SEQ ID NO: 2; preferably saidendoglucanase variant has activity on xanthan gum pretreated withxanthan lyase, further preferably said activity is a xanthan gumdegrading activity.

2. The detergent composition comprising an endoglucanase variant ofparagraph 1, which is a variant of a parent endoglucanase selected fromthe group of:

-   -   a) a polypeptide having at least about 60% sequence identity to        the mature polypeptide of SEQ ID NO: 2;    -   b) a polypeptide encoded by a polynucleotide that hybridizes        under low stringency conditions with (i) the mature polypeptide        coding sequence of SEQ ID NO: 1, or (ii) the full-length        complement of (i);    -   c) a polypeptide encoded by a polynucleotide having at least        about 60% identity to the mature polypeptide coding sequence of        SEQ ID NO: 1; and

d) a fragment of the mature polypeptide of SEQ ID NO: 2, which hasendoglucanase activity.

3. The detergent composition comprising an endoglucanase variant ofparagraph 2, wherein the parent endoglucanase having at least about 60%,e.g., at least about 65%, at least about 70%, at least about 75%, atleast about 80%, at least about 85%, at least about 90%, at least about95%, at least about 96%, at least about 97%, at least about 98%, atleast about 99% or about 100% sequence identity to the maturepolypeptide of SEQ ID NO: 2.

4. The detergent composition comprising an endoglucanase variant of anyof paragraphs 2-3, wherein the parent endoglucanase is encoded by apolynucleotide that hybridizes under low stringency conditions, mediumstringency conditions, medium-high stringency conditions, highstringency conditions, or very high stringency conditions with (i) themature polypeptide coding sequence of SEQ ID NO: 1 or (ii) thefull-length complement of (i).

5. The detergent composition comprising an endoglucanase variant of anyof paragraphs 2-4, wherein the parent endoglucanase is encoded by apolynucleotide having at least about 60%, e.g., at least about 65%, atleast about 70%, at least about 75%, at least about 80%, at least about85%, at least about 90%, at least about 95%, at least about 96%, atleast about 97%, at least about 98%, at least about 99%, or about 100%sequence identity to the mature polypeptide coding sequence of SEQ IDNO: 1.

6. The detergent composition comprising an endoglucanase variant of anyof paragraphs 2-5, wherein the parent endoglucanase comprises orconsists of the mature polypeptide of SEQ ID NO: 2.

7. The detergent composition comprising an endoglucanase variant of anyof paragraphs 2-6, wherein the parent endoglucanase is a fragment of themature polypeptide of SEQ ID NO: 2, wherein the fragment hasendoglucanase activity.

8. The detergent composition comprising an endoglucanase variant of anyof paragraphs 2-7, which has at least about 60%, e.g., at least about65%, at least about 70%, at least about 75%, at least about 80%, atleast about 85%, at least about 90%, at least about 95% identity, atleast about 96%, at least about 97%, at least about 98%, or at leastabout 99%, but less than about 100%, sequence identity to the amino acidsequence of the parent endoglucanase.

9. The detergent composition comprising an endoglucanase variant of anyof paragraphs 1-8, wherein said region selected from the group ofregions 1-9 is a chelator-induced instability region;

preferably said chelator-induced instability region (e.g., of SEQ ID NO:2 or another parent endoglucanase) has one or more of the followingfeatures:

i) in the presence of a chelator it is less conformationally stable thanone or more or all of its adjacent regions; and/or

ii) in the presence of a chelator it is more exposed to said chelatorthan one or more or all of its adjacent regions; and/or

iii) in the presence of a chelator it is more accessible to saidchelator than one or more or all of its adjacent regions; and/or

iv) in the presence of a chelator it is more conformationally dynamicthan one or more or all of its adjacent regions; and/or

v) in the presence of a chelator it is more receptive to deuteriumincorporation than one or more or all of its adjacent regions;

further preferably said adjacent region is selected from the group of:

-   -   vi) region 10 corresponding to amino acids 1 to 94 of SEQ ID NO:        2,    -   vii) region 11 corresponding to amino acids 106 to 114 of SEQ ID        NO: 2,    -   viii) region 12 corresponding to amino acids 139 to 209 of SEQ        ID NO: 2,    -   ix) region 13 corresponding to amino acids 252 to 266 of SEQ ID        NO: 2,    -   x) region 14 corresponding to amino acids 302 to 338 of SEQ ID        NO: 2,    -   xi) region 15 corresponding to amino acids 362 to 546 of SEQ ID        NO: 2,    -   xii) region 16 corresponding to amino acids 596 to 611 of SEQ ID        NO: 2,    -   xiii) region 17 corresponding to amino acids 661 to 805 of SEQ        ID NO: 2,    -   xiv) region 18 corresponding to amino acids 829 to 838 of SEQ ID        NO: 2, and    -   xv) region 19 corresponding to amino acids 1043 to 1055 of SEQ        ID NO: 2;

further most preferably said chelator is EDTA or citrate.

10. The detergent composition comprising an endoglucanase variant of anyone of paragraphs 1-9, wherein said variant further comprises analteration in at least one adjacent region, said adjacent region isselected from the group of:

-   -   i′) region 10 corresponding to amino acids 1 to 94 of SEQ ID NO:        2,    -   ii′) region 11 corresponding to amino acids 106 to 114 of SEQ ID        NO: 2,    -   iii′) region 12 corresponding to amino acids 139 to 209 of SEQ        ID NO: 2,    -   iv′) region 13 corresponding to amino acids 252 to 266 of SEQ ID        NO: 2,    -   v′) region 14 corresponding to amino acids 302 to 338 of SEQ ID        NO: 2,    -   vi′) region 15 corresponding to amino acids 362 to 546 of SEQ ID        NO: 2,    -   vii′) region 16 corresponding to amino acids 596 to 611 of SEQ        ID NO: 2,    -   viii′) region 17 corresponding to amino acids 661 to 805 of SEQ        ID NO: 2,    -   ix′) region 18 corresponding to amino acids 829 to 838 of SEQ ID        NO: 2, and    -   x′) region 19 corresponding to amino acids 1043 to 1055 of SEQ        ID NO: 2.

11. The detergent composition comprising an endoglucanase variant of anyof paragraphs 1-10, wherein in an aqueous solution comprising adetergent component said region (e.g., of SEQ ID NO: 2 or another parentendoglucanase) selected from the group of regions 1-9 is lessconformationally stable than one or more or all of its adjacent regions;

preferably said adjacent region is selected from the group of:

-   -   i) region 10 corresponding to amino acids 1 to 94 of SEQ ID NO:        2,    -   ii) region 11 corresponding to amino acids 106 to 114 of SEQ ID        NO: 2,    -   iii) region 12 corresponding to amino acids 139 to 209 of SEQ ID        NO: 2,    -   iv) region 13 corresponding to amino acids 252 to 266 of SEQ ID        NO: 2,    -   v) region 14 corresponding to amino acids 302 to 338 of SEQ ID        NO: 2,    -   vi) region 15 corresponding to amino acids 362 to 546 of SEQ ID        NO: 2,    -   vii) region 16 corresponding to amino acids 596 to 611 of SEQ ID        NO: 2,    -   viii) region 17 corresponding to amino acids 661 to 805 of SEQ        ID NO: 2,    -   ix) region 18 corresponding to amino acids 829 to 838 of SEQ ID        NO: 2, and    -   x) region 19 corresponding to amino acids 1043 to 1055 of SEQ ID        NO: 2;

further preferably said detergent component comprises a chelator;further most preferably said chelator is EDTA or citrate.

12. The detergent composition comprising an endoglucanase variant of anyof paragraphs 1-11, wherein in an aqueous solution comprising adetergent component said region (e.g., of SEQ ID NO: 2 or another parentendoglucanase) selected from the group of regions 1-9 is more exposed tosaid detergent component than one or more or all of its adjacentregions;

preferably said adjacent region is selected from the group of:

-   -   i) region 10 corresponding to amino acids 1 to 94 of SEQ ID NO:        2,    -   ii) region 11 corresponding to amino acids 106 to 114 of SEQ ID        NO: 2,    -   iii) region 12 corresponding to amino acids 139 to 209 of SEQ ID        NO: 2,    -   iv) region 13 corresponding to amino acids 252 to 266 of SEQ ID        NO: 2,    -   v) region 14 corresponding to amino acids 302 to 338 of SEQ ID        NO: 2,    -   vi) region 15 corresponding to amino acids 362 to 546 of SEQ ID        NO: 2,    -   vii) region 16 corresponding to amino acids 596 to 611 of SEQ ID        NO: 2,    -   viii) region 17 corresponding to amino acids 661 to 805 of SEQ        ID NO: 2,    -   ix) region 18 corresponding to amino acids 829 to 838 of SEQ ID        NO: 2, and    -   x) region 19 corresponding to amino acids 1043 to 1055 of SEQ ID        NO: 2;

further preferably said detergent component comprises a chelator;further most preferably said chelator is EDTA or citrate.

13. The detergent composition comprising an endoglucanase variant of anyof paragraphs 1-12, wherein in an aqueous solution comprising adetergent component said region (e.g., of SEQ ID NO: 2 or another parentendoglucanase) selected from the group of regions 1-9 is more accessibleto said detergent component than one or more or all of its adjacentregions;

preferably said adjacent region is selected from the group of:

-   -   i) region 10 corresponding to amino acids 1 to 94 of SEQ ID NO:        2,    -   ii) region 11 corresponding to amino acids 106 to 114 of SEQ ID        NO: 2,    -   iii) region 12 corresponding to amino acids 139 to 209 of SEQ ID        NO: 2,    -   iv) region 13 corresponding to amino acids 252 to 266 of SEQ ID        NO: 2,    -   v) region 14 corresponding to amino acids 302 to 338 of SEQ ID        NO: 2,    -   vi) region 15 corresponding to amino acids 362 to 546 of SEQ ID        NO: 2,    -   vii) region 16 corresponding to amino acids 596 to 611 of SEQ ID        NO: 2,    -   viii) region 17 corresponding to amino acids 661 to 805 of SEQ        ID NO: 2,    -   ix) region 18 corresponding to amino acids 829 to 838 of SEQ ID        NO: 2, and    -   x) region 19 corresponding to amino acids 1043 to 1055 of SEQ ID        NO: 2;

further preferably said detergent component comprises a chelator;further most preferably said chelator is the EDTA or citrate.

14. The detergent composition comprising an endoglucanase variant of anyof paragraphs 1-13, wherein in an aqueous solution comprising adetergent component said region (e.g., of SEQ ID NO: 2 or another parentendoglucanase) selected from the group including of regions 1-9 is moreconformationally dynamic than one or more or all of its adjacentregions;

preferably said adjacent region is selected from the group of:

-   -   i) region 10 corresponding to amino acids 1 to 94 of SEQ ID NO:        2,    -   ii) region 11 corresponding to amino acids 106 to 114 of SEQ ID        NO: 2,    -   iii) region 12 corresponding to amino acids 139 to 209 of SEQ ID        NO: 2,    -   iv) region 13 corresponding to amino acids 252 to 266 of SEQ ID        NO: 2,    -   v) region 14 corresponding to amino acids 302 to 338 of SEQ ID        NO: 2,    -   vi) region 15 corresponding to amino acids 362 to 546 of SEQ ID        NO: 2,    -   vii) region 16 corresponding to amino acids 596 to 611 of SEQ ID        NO: 2,    -   viii) region 17 corresponding to amino acids 661 to 805 of SEQ        ID NO: 2,    -   ix) region 18 corresponding to amino acids 829 to 838 of SEQ ID        NO: 2, and    -   x) region 19 corresponding to amino acids 1043 to 1055 of SEQ ID        NO: 2;

further preferably said detergent component comprises a chelator;further most preferably said chelator is EDTA or citrate.

15. The detergent composition comprising an endoglucanase variant of anyof paragraphs 1-14, wherein in an aqueous solution comprising adetergent component said region (e.g., of SEQ ID NO: 2 or another parentendoglucanase) selected from the group of regions 1-9 is more receptiveto deuterium incorporation than one or more or all of its adjacentregions;

preferably said adjacent region is selected from the group of:

-   -   i) region 10 corresponding to amino acids 1 to 94 of SEQ ID NO:        2,    -   ii) region 11 corresponding to amino acids 106 to 114 of SEQ ID        NO: 2,    -   iii) region 12 corresponding to amino acids 139 to 209 of SEQ ID        NO: 2,    -   iv) region 13 corresponding to amino acids 252 to 266 of SEQ ID        NO: 2,    -   v) region 14 corresponding to amino acids 302 to 338 of SEQ ID        NO: 2,    -   vi) region 15 corresponding to amino acids 362 to 546 of SEQ ID        NO: 2,    -   vii) region 16 corresponding to amino acids 596 to 611 of SEQ ID        NO: 2,    -   viii) region 17 corresponding to amino acids 661 to 805 of SEQ        ID NO: 2,    -   ix) region 18 corresponding to amino acids 829 to 838 of SEQ ID        NO: 2, and    -   x) region 19 corresponding to amino acids 1043 to 1055 of SEQ ID        NO: 2;

further preferably said detergent component comprises a chelator;further most preferably said chelator is EDTA or citrate.

16. The detergent composition comprising an endoglucanase variant of anyof paragraphs 1-15, further comprising an alteration (e.g., asubstitution, deletion or insertion) at one or more positions in:

a) one or more regions selected from the group of:

-   -   i) region 1 corresponding to amino acids 95 to 105 of SEQ ID NO:        2, e.g., said alteration at one or more positions selected from        the group of positions: 95, 96, 97, 98, 99, 100, 101, 102, 103,        104, 105, wherein said positions correspond to amino acid        positions of SEQ ID NO: 2 (e.g., using the numbering of SEQ ID        NO: 2),    -   ii) region 2 corresponding to amino acids 115 to 138 of SEQ ID        NO: 2, e.g., said alteration at one or more positions selected        from the group of positions: 115, 116, 117, 118, 119, 120, 121,        122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134,        135, 136, 137, 138, wherein said positions correspond to amino        acid positions of SEQ ID NO: 2 (e.g., using the numbering of SEQ        ID NO: 2),    -   iii) region 3 corresponding to amino acids 210 to 251 of SEQ ID        NO: 2, e.g., said alteration at one or more positions selected        from the group of positions: 210, 211, 212, 213, 214, 215, 216,        217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229,        230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 242,        243, 244, 245, 246, 247, 248, 249, 250, 251, wherein said        positions correspond to amino acid positions of SEQ ID NO: 2        (e.g., using the numbering of SEQ ID NO: 2),    -   iv) region 4 corresponding to amino acids 267 to 301 of SEQ ID        NO: 2, e.g., said alteration at one or more positions selected        from the group of positions: 267, 268, 269, 270, 271, 272, 273,        274, 275, 276, 277, 278, 279, 280, 281, 282, 283, 284, 285, 286,        287, 288, 289, 290, 291, 292, 293, 294, 295, 296, 297, 298, 299,        300, 301, wherein said positions correspond to amino acid        positions of SEQ ID NO: 2 (e.g., using the numbering of SEQ ID        NO: 2),    -   v) region 5 corresponding to amino acids 339 to 361 of SEQ ID        NO: 2, e.g., said alteration at one or more positions selected        from the group of positions: 339, 340, 341, 342, 343, 344, 345,        346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357, 358,        359, 360, 361, wherein said positions correspond to amino acid        positions of SEQ ID NO: 2 (e.g., using the numbering of SEQ ID        NO: 2),    -   vi) region 6 corresponding to amino acids 547 to 595 of SEQ ID        NO: 2, e.g., said alteration at one or more positions selected        from the group of positions: 547, 548, 549, 550, 551, 552, 553,        554, 555, 556, 557, 558, 559, 560, 561, 562, 563, 564, 565, 566,        567, 568, 569, 570, 571, 572, 573, 574, 575, 576, 577, 578, 579,        580, 581, 582, 583, 584, 585, 586, 587, 588, 589, 590, 591, 592,        593, 594, 595, wherein said positions correspond to amino acid        positions of SEQ ID NO: 2 (e.g., using the numbering of SEQ ID        NO: 2),    -   vii) region 7 corresponding to amino acids 612 to 660 of SEQ ID        NO: 2, e.g., said alteration at one or more positions selected        from the group of positions: 612, 613, 614, 615, 616, 617, 618,        619, 620, 621, 622, 623, 624, 625, 626, 627, 628, 629, 630, 631,        632, 633, 634, 635, 636, 637, 638, 639, 640, 641, 642, 643, 644,        645, 646, 647, 648, 649, 650, 651, 652, 653, 654, 655, 656, 657,        658, 659, 660, wherein said positions correspond to amino acid        positions of SEQ ID NO: 2 (e.g., using the numbering of SEQ ID        NO: 2),    -   viii) region 8 corresponding to amino acids 806 to 828 of SEQ ID        NO: 2, e.g., said alteration at one or more positions selected        from the group of positions: 806, 807, 808, 809, 810, 811, 812,        813, 814, 815, 816, 817, 818, 819, 820, 821, 822, 823, 824, 825,        826, 827, 828, wherein said positions correspond to amino acid        positions of SEQ ID NO: 2 (e.g., using the numbering of SEQ ID        NO: 2), and    -   ix) region 9 corresponding to amino acids 839 to 1042 of SEQ ID        NO: 2, e.g., said alteration at one or more positions selected        from the group of positions: 839, 840, 841, 842, 843, 844, 845,        846, 847, 848, 849, 850, 851, 852, 853, 854, 855, 856, 857, 858,        859, 860, 861, 862, 863, 864, 865, 866, 867, 868, 869, 870, 871,        872, 873, 874, 875, 876, 877, 878, 879, 880, 881, 882, 883, 884,        885, 886, 887, 888, 889, 890, 891, 892, 893, 894, 895, 896, 897,        898, 899, 900, 901, 902, 903, 904, 905, 906, 907, 908, 909, 910,        911, 912, 913, 914, 915, 916, 917, 918, 919, 920, 921, 922, 923,        924, 925, 926, 927, 928, 929, 930, 931, 932, 933, 934, 935, 936,        937, 938, 939, 940, 941, 942, 943, 944, 945, 946, 947, 948, 949,        950, 951, 952, 953, 954, 955, 956, 957, 958, 959, 960, 961, 962,        963, 964, 965, 966, 967, 968, 969, 970, 971, 972, 973, 974, 975,        976, 977, 978, 979, 980, 981, 982, 983, 984, 985, 986, 987, 988,        989, 990, 991, 992, 993, 994, 995, 996, 997, 998, 999, 1000,        1001, 1002, 1003, 1004, 1005, 1006, 1007, 1008, 1009, 1010,        1011, 1012, 1013, 1014, 1015, 1016, 1017, 1018, 1019, 1020,        1021, 1022, 1023, 1024, 1025, 1026, 1027, 1028, 1029, 1030,        1031, 1032, 1033, 1034, 1035, 1036, 1037, 1038, 1039, 1040,        1041, 1042, wherein said positions correspond to amino acid        positions of SEQ ID NO: 2 (e.g., using the numbering of SEQ ID        NO: 2); and/or

b) an adjacent region (e.g., an alteration at one or more positionscorresponding to positions: 51 (e.g, K51Q), 451 (e.g., K451S), 333(e.g., W333L), 416 (e.g., Q416D)), preferably said adjacent region isselected from the group of:

-   -   i′) region 10 corresponding to amino acids 1 to 94 of SEQ ID NO:        2,    -   ii′) region 11 corresponding to amino acids 106 to 114 of SEQ ID        NO: 2,    -   iii′) region 12 corresponding to amino acids 139 to 209 of SEQ        ID NO: 2,    -   iv′) region 13 corresponding to amino acids 252 to 266 of SEQ ID        NO: 2,    -   v′) region 14 corresponding to amino acids 302 to 338 of SEQ ID        NO: 2,    -   vi′) region 15 corresponding to amino acids 362 to 546 of SEQ ID        NO: 2,    -   vii′) region 16 corresponding to amino acids 596 to 611 of SEQ        ID NO: 2,    -   viii′) region 17 corresponding to amino acids 661 to 805 of SEQ        ID NO: 2,    -   ix′) region 18 corresponding to amino acids 829 to 838 of SEQ ID        NO: 2, and    -   x′) region 19 corresponding to amino acids 1043 to 1055 of SEQ        ID NO: 2,

wherein said variant has at least about 60%%, e.g., at least about 65%,at least about 70%, at least about 75%, at least about 80%, at leastabout 85%, at least about 90%, at least about 95%, at least about 96%,at least about 97%, at least about 98%, or at least about 99%, and lessthan about 100% sequence identity to SEQ ID NO: 2, preferably saidvariant has activity on xanthan gum pretreated with xanthan lyase,further preferably said activity is a xanthan gum degrading activity.

17. The detergent composition comprising an endoglucanase variant of anyof paragraphs 1-16, wherein said variant has at least about 61%, atleast about 62%, at least about 63%, at least about 64%, at least about65%, at least about 66%, at least about 67%, at least about 68%, atleast about 69%, at least about 70%, at least about 71%, at least about72%, at least about 73%, at least about 74%, at least about 75%, atleast about 76%, at least about 77%, at least about 78%, at least about79%, at least about 80%, at least about 81%, at least about 82%, atleast about 83%, at least about 84%, at least about 85%, at least about86%, at least about 87%, at least about 88%, at least about 89%, atleast about 90%, at least about 91%, at least about 92%, at least about93%, at least about 94%, at least about 95%, at least about 96%, atleast about 97%, at least about 98%, or at least about 99% sequenceidentity to SEQ ID NO: 2.

18. The detergent composition comprising an endoglucanase variant of anyone of paragraphs 1-17, wherein said alteration at one or more positionis selected from the group of alterations in positions: 4, 17, 18, 20,51, 53, 55, 56, 60, 63, 71, 79, 87, 92, 99, 120, 125, 126, 130, 137,182, 186, 189, 192, 213, 216, 221, 226, 228, 230, 231, 232, 233, 235,240, 243, 247, 249, 278, 279, 281, 283, 285, 289, 292, 294, 298, 302,311, 313, 333, 346, 353, 358, 386, 387, 388, 390, 403, 408, 410, 416,441, 448, 451, 471, 472, 476, 489, 507, 512, 515, 538, 555, 556, 557,558, 559, 560, 561, 562, 563, 564, 567, 568, 570, 575, 578, 579, 580,581, 583, 589, 590, 591, 592, 593, 595, 598, 599, 602, 603, 605, 607,609, 616, 627, 630, 631, 635, 636, 638, 639, 640, 641, 642, 643, 644,651, 676, 683, 688, 690, 694, 698, 699, 706, 711, 713, 1719, 720, 744,749, 754, 756, 760, 781, 786, 797, 810, 811, 812, 815, 823, 824, 825,827, 828, 833, 834, 835, 837, 843, 848, 868, 869, 870, 871, 872, 873,874, 880, 881, 883, 884, 885, 887, 888, 890, 892, 894, 898, 905, 906,912, 920, 921, 924, 926, 927, 928, 932, 933, 934, 935, 937, 938, 939,940, 941, 942, 943, 946, 948, 950, 952, 953, 954, 956, 957, 960, 966,971, 972, 980, 989, 991, 994, 995, 998, 999, 1006, 1009, 1010, 1011,1029, 1030, 1031, 1032, 1035, 1037, 1038, 1040, 1041, 1042, 1044, 1045,1048, wherein numbering is according to SEQ ID NO: 2.

19. The detergent composition comprising an endoglucanase variant of anyof paragraphs 1-18, wherein said alteration at one or more positions isselected from the group of alterations in positions: 285, 333, 353,*558, 558, 633, 635, 635, 635, 638, 639, 994, 281, 563, 575, 575, 921,558+559+560+561+562, 558, 559, 560, 561, 562 125, 126, 130, 213, 221,228, 228, 230, 230, 230, 230, 230, 230, 230, 231, 231, 232, 232, 235,240, 243, 243, 249, 278, 281, 281, 281, 281, 281, 281, 281, 285, 285,285, 285, 285, 285, 285, 285, 285, 285, 285, 292, 292, 292, 292, 292,292, 292, 292, 292, 292, 292, 292, 297, 346, 556, 558, 558, 558, 558,558, 558, 559, 559, 559, 559, 559, 559, 559, 559, 559, 559, 559, 559,560, 560, 560, 560, 561, 561, 561, 561, 561, 564, 564, 564, 564, 564,564, 565, 567, 568, 569, 569, 569, 569, 570, 570, 570, 570, 570, 570,570, 570, 570, 570, 570, 570, 570, 570, 570, 570, 570, 575, 575, 576,576, 576, 578, 579, 579, 580, 583, 589, 590, 590, 590, 591, 592, 593,593, 593, 593, 593, 593, 593, 593, 616, 627, 627, 627, 627, 627, 627,627, 630, 630, 630, 635, 635, 635, 635, 635, 635, 635, 636, 636, 636,636, 636, 636, 636, 636, 638, 638, 638, 638, 638, 639, 639, 639, 639,639, 639, 639, 639, 639, 641, 642, 642, 643, 643, 643, 644, 651, 810,811, 812, 812, 812, 812, 812, 815, 815, 815, 815, 815, 823, 824, 825,825, 825, 825, 827, 827, 827, 843, 870, 870, 870, 870, 870, 870, 870,870, 870, 870, 870, 870, 870, 870, 870, 870, 871, 871, 871, 871, 871,871, 871, 871, 871, 871, 871, 872, 872, 872, 872, 872, 872, 872, 872,872, 872, 872, 872, 872, 873, 873, 874, 874, 874, 874, 874, 874, 874,874, 881, 883, 884, 885, 885, 885, 887, 887, 887, 887, 887, 894, 920,921, 921, 932, 933, 933, 934, 934, 934, 934, 934, 934, 934, 934, 934,935, 937, 937, 937, 937, 937, 937, 937, 938, 939, 939, 940, 941, 941,941, 942, 942, 943, 943, 950, 950, 950, 952, 952, 953, 954, 960, 964,964, 966, 966, 971, 974, 974, 989, 991, 991, 991, 991, 991, 991, 991,995, 995, 995, 995, 995, 998, 998, 1006, 1006, 1006, 1006, 1010, 1011,1011, 1011, 1011, 1011, 1011, 1011, 1029, 1030, 1031, 1031, 1031, 1031,1032, 1035, 1037, 1037, 1038, 1038, 1040, 1040, 1041, 1044, 1044, 1044,1044, 1045, 1045, 559+579, 559, 579, 564+579, 564, 579, 559+579, 559,579, 562+579, 562, 579, 564+579, 564, 579, 559+579+99, 559, 579, 99,559+579+281, 559, 579, 281, 281+559+579, 281, 559, 579, 559+579+616,559, 579, 616, 559+579+636, 559, 579, 636, 559+579+651, 559, 579, 651,559+579+948, 559, 579, 948, 559+579+1009, 559, 579, 1009, 559+579+627,559, 579, 627, 579+921, 579, 921, 559+579+921, 559, 579, 921, 99+579,99, 579, 579+651, 579, 651, 579+948, 579, 948, 579+1009, 579, 1009,559+579+934, 559, 579, 934, 559+579+921+934, 559, 579, 921, 934,559+579+627, 559, 579, 627, 559+579+627+616, 559, 579, 627, 616,559+579+627, 559, 579, 627, 559+579+921+651, 559, 579, 921, 651,559+579+921+627, 559, 579, 921, 627, 559+579+921+636, 559, 579, 921,636, 559+579+921+616, 559, 579, 921, 616, 559+579+921+636, 559, 579,921, 636, 559+579+921+627+636, 559, 579, 921, 627, 636, 559+579+636+651,559, 579, 636, 651, 559+579+616+651, 559, 579, 616, 651,559+579+616+636, 559, 579, 616, 636, 559+579+616+921+934, 559, 579, 616,921, 934, 559+579+651+627, 559, 579, 651, 627, 559+579+651+636, 559,579, 651, 636, 559+579+651+627+636, 559, 579, 651, 627, 636,559+579+651+616, 559, 579, 651, 616, 559+579+651+921+934, 559, 579, 651,921, 934, 636+934, 636, 934, 636+921, 636, 921, 636+627, 636, 627,636+579, 636, 579, 638+934, 638, 934, 638+921, 638, 921, 638+627, 638,627, 638+579, 638, 579, 627+51, 627, 51, 627+451, 627, 451, 627+559,627, 559, 627+579, 627, 579, 579+934, 579, 934, 651+638, 651, 638,570+651, 570, 651, 570+921, 570, 921, 570+627, 570, 627, 570+559, 570,559, 570+579, 570, 579, 570+638, 570, 638, 570+579, 570, 579, 570+638,570, 638, 570+651, 570, 651, 570+636, 570, 636, 570+934, 570, 934,570+638, 570, 638, 570+921, 570, 921, 570+627, 570, 627, 570+559, 570,559, 570+885, 570, 885, 885+934, 885, 934, 885+627, 885, 627,559+579+636, 559, 579, 636, 559+579+638, 559, 579, 638, 559+579+870,870, 559+579+560, 560, 559+579+564, 564, 559+579+570, 570, 559+579+570,570, 559+579+570, 570, 559+579+570, 570, 559+579+570, 570, 559+579+570,570, 559+579+570, 570, 559+579+570, 570, 558, 559, 559, 559, 561, 564,570, 570, 570, 570, 570, 570, 570, 579, 579, 581, 616, 627, 627, 627,636, 636, 636, 636, 636, 636, 638, 638, 643, 651, 651, 885, 885, 921,934, 934, 966, 1011, 1031, 559+570+579, 559+570+579, 559+570+579,559+570+579, 559+570+579, 559+570+579, 559+570+579, 559+570+579,559+560+579, 559+579+651, 559+579+651+934, 559+579+638, 559+579+921,559+579+616+921, 559+579+636, 559+579, 559+579, 559+579+921,559+579+616, 638+934, 627+636, 627+934, 570+579, 416+559+579+636, 416,128+559+579+627, 128, 128+559+579+636, 579+636 of SEQ ID NO: 2,preferably numbering is according to SEQ ID NO: 2, further preferablyalterations in positions: 627, 636 or 638, wherein numbering isaccording to SEQ ID NO: 2.

20. The detergent composition comprising an endoglucanase variant of anyof paragraphs 1-17, wherein said alteration at one or more positions isselected from the group of: N285G, W333L, T353D, N558NP, N558F, T633V,D635L, D635M, D635T, F638Y, T639D, G994N, and K281T, G563E, I575M,I575A, K921D, N558K+A559K+S560F+T561P+G562W, N558K, A559K, S560F, T561P,G562W and I125V, A126R, K130R, K213R, A221R, K228E, K228I, G230F, G230L,G230A, G230H, G230N, G230W, G230T, F231Y, F231N, V232R, V232G, H235D,N240Q, G243K, G243R, A249N, A278S, K281F, K281V, K281Y, K281H, K281Q,K281N, K281W, N285L, N285M, N285S, N285P, N285T, N285Y, N285H, N285K,N285D, N285W, N285R, T292F, T292L, T292I, T292V, T292S, T292P, T292Y,T292Q, T292N, T292K, T292D, T292G, F297L, A346H, G556S, N558D, N558M,N558Q, N558I, N558Y, N558H, A559N, A559F, A559M, A559P, A559Y, A559H,A559Q, A559D, A559R, A559G, A559I, A559S, S560P, S560K, S560G, S560D,T561P, T561E, T561Q, T561S, T561D, A564I, A564Y, A564H, A564Q, A564K,A564E, E565M, V567F, K568R, L569F, L569Y, L569D, L569E, P570F, P570L,P570I, P570M, P570V, P570S, P570T, P570A, P570Y, P570H, P570Q, P570N,P570K, P570E, P570W, P570R, P570G, I575D, I575E, I576F, I576M, I576P,D578R, Y579F, Y579W, V580L, D583M, Q589G, P590S, P590T, P590E, E591L,G592D, S593P, S593H, S593Q, S593N, S593K, S593D, S593E, S593R, S616D,K627L, K627M, K627V, K627S, K627T, K627Q, K627R, I630F, I630V, I630Y,D635A, D635P, D635N, D635K, D635E, D635G, D635W, S636L, S636M, S636A,S636H, S636Q, S636N, S636K, S636R, F638I, F638V, F638T, F638L, F638H,T639V, T639S, T639L, T639I, T639M, T639A, T639E, T639W, T639G, Y641E,S642T, S642N, N643D, N643H, N643T, T644F, A651P, S810R, A811S, V812F,V812I, V812M, V812W, V812R, N815V, N815Y, N815E, N815W, N815R, S823Q,A824T, T825N, T825W, T825A, T825D, V827I, V827M, V827S, T843V, D870F,D870L, D870I, D870M, D870V, D870S, D870T, D870Y, D870H, D870Q, D870N,D870K, D870E, D870W, D870R, D870G, P871F, P871L, P871I, P871M, P871V,P871S, P871T, P871A, P871Y, P871H, P871Q, T872S, T872F, T872A, T872Y,T872H, T872Q, T872N, T872K, T872D, T872E, T872W, T872R, T872G, D873K,D873E, T874V, T874S, T874P, T874A, T874H, T874Q, T874N, T874K, V881Q,T883K, Y884H, A885F, A885Q, A885N, T887L, T887I, T887S, T887H, T887R,K894E, N920D, K921R, K921E, T932A, N933V, N933S, Y934G, Y934M, Y934S,Y934A, Y934Q, Y934N, Y934E, Y934W, Y934R, T935W, A937F, A937V, A937S,A937T, A937Q, A937D, A937E, V938I, K939I, K939V, D940E, N941S, N941H,N941D, A942P, A942E, D943Y, D943H, R950V, R950H, R950N, F952S, F952W,N953Y, G954L, Y960F, A964N, A964C, N966P, N966C, G971A, Q974K, Q974C,Q989I, Q991L, Q991I, Q991M, Q991V, Q991T, Q991K, Q991C, S995I, S995V,S995Q, S995R, S995C, G998V, G998A, S1006T, S1006A, S1006K, S1006R,Y1010W, L1011M, L1011S, L1011A, L1011Q, L1011N, L1011D, L1011E, R1029N,F1030M, K1031I, K1031S, K1031T, K1031H, V1032G, K1035A, A1037E, A1037W,S1038L, S1038I, L1040N, L1040E, G1041F, L1044F, L1044S, L1044N, L1044W,P1045Q, P1045W, and A559N+Y579F, A559N, Y579F, A564E+Y579F, A564E,Y579F, A559N+Y579W, A559N, Y579W, G562P+Y579W, G562P, Y579W,A564D+Y579W, A564D, Y579W, A559N+Y579W+K99R, A559N, Y579W, K99R,A559N+Y579W+K281R, A559N, Y579W, K281R, K281R+A559N+Y579W, K281R, A559N,Y579W, A559N+Y579W+S616D, A559N, Y579W, S616D, A559N+Y579W+S636N, A559N,Y579W, S636N, A559N+Y579W+A651P, A559N, Y579W, A651P, A559N+Y579W+K948E,A559N, Y579W, K948E, A559N+Y579W+K1009E, A559N, Y579W, K1009E,A559N+Y579W+K627R, A559N, Y579W, K627R, Y579W+K921R, Y579W, K921R,A559N+Y579W+K921R, A559N, Y579W, K921R, K99R+Y579W, K99R, Y579W,Y579W+A651P, Y579W, A651P, Y579W+K948E, Y579W, K948E, Y579W+K1009E,Y579W, K1009E, A559N+Y579W+Y934G, A559N, Y579W, Y934G,A559N+Y579W+K921R+Y934G, A559N, Y579W, K921R, Y934G, A559N+Y579W+K627M,A559N, Y579W, K627M, A559N+Y579W+K627R+S616D, A559N, Y579W, K627R,S616D, A559N+Y579F+K627R, A559N, Y579F, K627R, A559N+Y579W+K921R+A651P,A559N, Y579W, K921R, A651P, A559N+Y579W+K921R+K627R, A559N, Y579W,K921R, K627R, A559N+Y579W+K921R+S636K, A559N, Y579W, K921R, S636K,A559N+Y579W+K921R+S616D, A559N, Y579W, K921R, S616D,A559N+Y579W+K921R+S636N, A559N, Y579W, K921R, S636N,A559N+Y579W+K921R+K627R+S636N, A559N, Y579W, K921R, K627R, S636N,A559N+Y579W+S636N+A651P, A559N, Y579W, S636N, A651P,A559N+Y579W+S616D+A651P, A559N, Y579W, S616D, A651P,A559N+Y579W+S616D+S636K, A559N, Y579W, S616D, S636K,A559N+Y579W+S616D+K921R+Y934G, A559N, Y579W, S616D, K921R, Y934G,A559N+Y579W+A651P+K627M, A559N, Y579W, A651P, K627M,A559N+Y579W+A651P+S636K, A559N, Y579W, A651P, S636K,A559N+Y579W+A651P+K627R+S636N, A559N, Y579W, A651P, K627R, S636N,A559N+Y579W+A651P+S616D, A559N, Y579W, A651P, S616D,A559N+Y579W+A651P+K921R+Y934G, A559N, Y579W, A651P, K921R, Y934G,S636N+Y934G, S636N, Y934G, S636N+K921R, S636N, K921R, S636N+K627R,S636N, K627R, S636N+Y579W, S636N, Y579W, F638I+Y934G, F638I, Y934G,F638I+K921R, F638I, K921R, F638I+K627R, F638I, K627R, F638I+Y579W,F638I, Y579W, K627R+K51Q, K627R, K51Q, K627R+K451S, K627R, K451S,K627R+A559N, K627R, A559N, K627R+Y579W, K627R, Y579W, Y579W+Y934G,Y579W, Y934G, A651P+F638I, A651P, F638I, P570Q+A651P, P570Q, A651P,P570Q+K921R, P570Q, K921R, P570Q+K627R, P570Q, K627R, P570Q+A559N,P570Q, A559N, P570Q+Y579W, P570Q, Y579W, P570Q+F638I, P570Q, F638I,P570K+Y579W, P570K, Y579W, P570K+F638I, P570K, F638I, P570T+A651P,P570T, A651P, P570T+S636N, P570T, S636N, P570T+Y934G, P570T, Y934G,P570T+F638I, P570T, F638I, P570T+K921R, P570T, K921R, P570T+K627R,P570T, K627R, P570T+A559N, P570T, A559N, P570T+A885F, P570T, A885F,A885F+Y934G, A885F, Y934G, A885F+K627R, A885F, K627R, A559N+Y579W+S636L,A559N, Y579W, S636L, A559N+Y579W+F638I, A559N, Y579W, F638I,A559N+Y579W+D870M, D870M, A559N+Y579W+S560P, S560P, A559N+Y579W+A564I,A564I, A559N+Y579W+P570N, P570N, A559N+Y579W+P570K, P570K,A559N+Y579W+P570R, P570R, A559N+Y579W+P570A, P570A, A559N+Y579W+P570T,P570T, A559N+Y579W+P570S, P570S, A559N+Y579W+P570Q, P570Q,A559N+Y579W+P570H, P570H, and N558E, A559P, A559N, A559H, T561P, A564E,P570A, P570Q, P570R, P570S, P570K, P570T, P570N, Y579W, Y579F, T581M,S616D, K627R, K627M, K627Q, S636N, S636Q, S636R, S636K, S636M, S636H,F638I, F638L, N643D, A651P, A651S, A885F, A885Q, K921R, Y934R, Y934G,N966C, L1011A, K1031I, and A559N+P570A+Y579W, A559N+P570H+Y579W,A559N+P570K+Y579W, A559N+P570N+Y579W, A559N+P570Q+Y579W,A559N+P570R+Y579W, A559N+P570S+Y579W, A559N+P570T+Y579W,A559N+S560P+Y579W, A559N+Y579W+A651P, A559N+Y579W+A651P+Y934G,A559N+Y579W+F638I, A559N+Y579W+K921R, A559N+Y579W+S616D+K921R,A559N+Y579W+S636N, A559N+Y579F, A559N+Y579W, A559N+Y579W+K921R,A559N+Y579W+S616D, F638I+Y934G, K627R+S636N, K627R+Y934G, P570K+Y579W,Q416D+A559N+Y579W+S636N, Q416D, S128X+A559N+Y579W+K627R, S128X,S128X+A559N+Y579W+S636N, Y579W+S636N, V4T, S17A, N18G, F20P, F20N, F20G,F20Y, K51Q, K51H, E53Y, E53P, E53G, Y55M, Y55D, V56M, Y60F, S63F, A71E,579W, T87R, T92S, A120P, N129D, F137L, H182Y, A186P, N189K, K192N,N216D, N216Q, N216R, L226K, G230H, L233H, D247N, G279E, K281R, A283D,N285D, N285G, Q289E, T292A, T292F, T292Y, A294V, Q298E, I302D, I302H,I302V, I302M, H311N, 5313D, A346D, A386P, I387T, K388R, K390Q, I403Y,E408D, E408N, E408S, E408P, E408A, E408G, P410G, Q416S, Q416D, N441G,A448E, A448W, A448S, K451S, K451Q, G471S, S472Y, D476R, Q489P, K507R,K512P, S515V, S538C, L555Q, G557R, N558E, A559N, A559P, A559H, A559D,S560P, S560G, T561P, A564E, A564I, V567P, K568R, P570R, P570Q, P570K,P570A, P570T, P570G, P570S, P570H, P570N, 1575V, Y579W, Y579F, T581M,S593N, S593E, S595L, S598Q, A599S, 1602T, 1602D, V603P, S605T, S607C,G609E, S616G, S616D, K627R, K627M, K627Q, K631R, K631A, D635A, D635E,D635M, D635N, D635L, D635W, S636N, S636K, S636L, S636Q, S636R, S636M,S636H, F638N, F638I, F638L, F638V, F638H, F638M, T639G, T639I, T639M,T639Y, T639W, T639P, T639E, T640S, S642N, S642T, N643D, N643H, A651P,A651S, D676H, Q683E, A688G, Y690F, T694A, T697G, R698W, T699A, T706Q,T711S, T711V, T711Y, K713R, W719R, K720H, K744H, K744Q, A749T, K754R,V756Y, V756H, S760G, T781M, N786K, T797S, S810Q, A824D, T825G, N828D,N833D, Q834E, S835A, S835D, V837I, N848D, A868E, A869V, D870V, T872G,T872H, T872W, T872Q, R880K, V881Q, V881T, T883R, T883V, T883C, T883K,Y884H, A885N, A885Q, A885F, T887K, T887S, L888M, V890R, T892P, T892V,R898Q, N905D, F906A, Q912V, N920P, K921R, A924D, V926F, V926P, K927R,S928D, T932A, N933S, N933V, Y934G, Y934R, Y934Q, A937E, V938I, K939V,N941S, A942P, G946R, K948R, Q956Y, Q956S, A957L, A957P, N966C, T972K,M980I, G994D, T999R, L1011A, K1031I, A1037E, S1038G, G1041R, Y1042N,F1048W, preferably numbering is according to SEQ ID NO: 2, furtherpreferably alterations in positions: K627R, S636N or F638I, whereinnumbering is according to SEQ ID NO: 2.

21. The detergent composition comprising an endoglucanase variant of anyof paragraphs 1-20, wherein the total number of alterations compared tothe parent endoglucanase (e.g., SEQ ID NO: 2) is between about 1 andabout 20, e.g., between about 1 and about 10 or between about 1 andabout 5, such as about 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 alterations.

22. The detergent composition comprising an endoglucanase variant of anyof paragraphs 1-21, wherein said activity on xanthan gum pretreated withxanthan lyase is a xanthan degrading activity, preferably said xanthandegrading activity is endoglucanase EC 3.2.1.4 activity.

23. The detergent composition comprising an endoglucanase variant of anyof paragraphs 1-22, wherein said variant has an improved stability in adetergent composition compared to a parent endoglucanase (e.g., with SEQID NO: 2); preferably said detergent composition comprises a chelator;further preferably said chelator is EDTA or citrate.

24. The detergent composition comprising an endoglucanase variant of anyof paragraphs 1-23, wherein said variant has a half-life improvementfactor (HIF) of ≥about 1.0; preferably said variant has a half-lifeimprovement factor (HIF) of ≥about 1.0.

25. The detergent composition comprising an endoglucanase variant ofparagraph 24, wherein said half-life improvement factor (HIF) isdetermined after incubation of said endoglucanase variant in a detergentcomposition at about 25° C. for a time period from about 17 to about 20hours.

26. The detergent composition of any of paragraphs 1-25, furthercomprising one or more components; preferably said component comprises achelator; further preferably said chelator is EDTA or citrate.

27. The detergent composition of any of paragraphs 1-26, furthercomprising one or more additional enzymes selected from the group of:endoglucanases, proteases, amylases, lichenases, lipases, cutinases,cellulases, xanthan lyases, xyloglucanases, pectinases, pectin lyases,xanthanases, peroxidases, haloperoxygenases, catalases and mannanases,or any mixture thereof.

28. The detergent composition of any of paragraphs 1-27, wherein saidcomposition is in form of a bar, a homogenous tablet, a tablet havingtwo or more layers, a pouch having one or more compartments, a regularor compact powder, a granule, a paste, a gel, or a regular, compact orconcentrated liquid.

29. Use of a detergent composition of any of paragraphs 1-28, whereinsaid use is for degrading xanthan gum.

30. The use of paragraph 29, wherein said endoglucanase variant has anenzyme detergency benefit.

The present disclosure is further described by the following examplesthat should not be construed as limiting the scope of the disclosure.

EXAMPLES Example 1: Hydrogen Exchange—HDX

Continuous amide Hydrogen/Deuterium (H/D) exchange of the parentendoglucanase of SEQ ID NO: 2 was initiated by addition of 99.9%deuterated 20 mM Tris, 1 mM CaCI2, pH 8 in the absence or presence of 10mM EDTA to a final deuterium concentration of 95%. H/D exchange wasperformed in triplicates at 22° C. at a concentration of 1 μM. At fivetime points ranging from 15 seconds to 1 hour the samples were quenchedby addition of 1:1 (v/v) ice-cold 6M guanidinium hydrochloride, 300 mMphosphate, pH 2.05 to a final pH of 2.6. The quenched samples wereimmediately frozen and stored at −80° C. until LC-MS analysis.Non-deuterated samples were prepared following the same procedure, butusing protiated buffers. Fully deuterated samples (95% D20 for theparent endoglucanase of SEQ ID NO: 2) were prepared by over-nightincubation in 99.9% deuterated 6M guanidinium hydrochloride and quenchedin 300 mM phosphate, pH 2.3 to a final pH of 2.6.

The quenched samples were loaded into a cooled HDX-UPLC system foronline pepsin digestion using an immobilized pepsin column (Pierce,Rockford, USA). The peptides were desalted using a trap column (WatersVanGuard C18, 1.7 μM, 2.1×5 mm) at a flow of 200 μl/min 0.23% formicacid for 3 min and peptides were separated by reverse phasechromatography (Waters Acquity BEH C18, 1.7 μm, 1×100 mm) using atwo-step gradient from 8-18% in 2 min and 18-40% in 10 min 0.23% formicacid in acetonitrile at a flow of 40 μl/min. Positive electrosprayionization mass spectrometry with ion mobility was performed on thepeptides using a Synapt G2 mass spectrometer (Waters, Milford, USA).

The peptic peptides of the mature parent endoglucanase of SEQ ID NO: 2were identified by tandem mass spectrometry of non-deuterated samplesusing a combination of data independent (MSe) and data dependentacquisition schemes (DDA) and data-analysis in Protein Lynx GlobalServer v. 2.5. Deuterium incorporation of individual peptides weredetermined in DynamX v. 3.0 or in HXexpress (M. Guttman, D. Weis, J.Engen, K. Lee, J. Am. Soc. Mass. Spectrom. 2013, 24, 1906-1912).Statistical analysis (F-tests and student T-tests) were employed todetermine statistically significant changes in H/D exchange between theanalyzed protein states.

Peptides with significant changes in H/D exchange (ΔDX>0.5) wereselected as identifying chelator-induced instability regions within SEQID NO: 2. The following chelator-induced instability regions wereidentified within SEQ ID NO: 2: region 1 corresponding to amino acids 95to 105 of SEQ ID NO: 2, region 2 corresponding to amino acids 115 to 138of SEQ ID NO: 2, region 3 corresponding to amino acids 210 to 251 of SEQID NO: 2, region 4 corresponding to amino acids 267 to 301 of SEQ ID NO:2, region 5 corresponding to amino acids 339 to 361 of SEQ ID NO: 2,region 6 corresponding to amino acids 547 to 595 of SEQ ID NO: 2, region7 corresponding to amino acids 612 to 660 of SEQ ID NO: 2, region 8corresponding to amino acids 806 to 828 of SEQ ID NO: 2, and region 9corresponding to amino acids 839 to 1042 of SEQ ID NO: 2.

Example 2: Construction of GH9 Endoglucanase Variants of the MatureParent Endoglucanase Having SEQ ID NO: 2

A linear integration vector-system was used for cloning of the matureparent nucleotide sequence having SEQ ID NO: 1 (same also disclosed asmature peptide within SEQ ID NO: 1 of WO2013167581A1) coding for themature parent polypeptide of the GH9 endoglucanase of SEQ ID NO: 2, andits variants. The linear integration construct was a PCR fusion made byfusing the gene between two Bacillus subtilis homologous chromosomalregions along with strong promoters and a chloramphenicol resistancemarker. The fusion was made by Splicing by Overlap Extension (SOE) PCR(Horton, R. M, Hunt, H. D., Ho, S. N., Pullen, J. K. and Pease, L. R.(1989). Engineering hybrid genes without the use of restriction enzymes,or gene splicing were produced by overlap extension (Gene 77: 61-68).The SOE PCR method is also described in patent application WO2003/095658. The gene was expressed under the control of a triplepromoter system (as described in WO 99/43835), including the promotersfrom Bacillus licheniformis alpha-amylase gene (amyL), Bacillusamyloliquefaciens alpha-amylase gene (amyQ), and the Bacillusthuringiensis crylllA promoter including stabilizing sequence. The genecoding for chloramphenicol acetyltransferase was used as marker(described in e.g. Diderichsen, B.; Poulsen, G. B.; Joergensen, S. T.; Auseful cloning vector for Bacillus subtilis. Plasmid 30:312, 1993). Thefinal gene constructs were integrated on the Bacillus chromosome byhomologous recombination into the pectate lyase locus. The genefragments were amplified from chromosomal DNA of the correspondingstrains with gene specific primers containing overhang to the twoflanking vector fragments. All genes were expressed with a Bacilluslicheniformis alpha-amylase secretion signal having the nucleotidesequence of SEQ ID NO: 3 and the amino acid sequence of SEQ ID NO: 4replacing the native secretion signal.

Variants of the mature parent GH9 endocluconase from Paenibacillussp-62047 having SEQ ID NO: 2 as described in Examples 3-4 below weremade by the megaprimer mutagenesis method using specifically designedmutagenic oligonucleotides introducing desired mutations in theresulting sequence. Design and production methods for such mutagenicoligonucleotides introducing desired mutations into target sequences arewell known to those skilled in the art. Consequently, mutagenic oligoswere designed and synthesized corresponding to the DNA sequence flankingthe desired site(s) of mutation, separated by the DNA base pairsdefining the substitutions. The final expression cassette composed thereference GH9 endocluconase from Paenibacillus sp-62047 as describedabove (i.e., parent GH9 endocluconase having SEQ ID NO: 1). Successfulintroduction of the desired substitutions was confirmed by DNAsequencing of the GH9 endogluconase gene. An aliquot of the PCR productwas subsequently transformed into Bacillus subtilis. Transformants wereselected on LB agar plates supplemented with 10 mM K2P04, 0.4% extraglucose and 6 μg of chloramphenicol per ml. The resulting recombinantBacillus subtilis clone containing the integrated expression constructwas grown in liquid culture as described below. The enzyme containingsupernatants were harvested and the enzymes (variants) were eitherstress tested using a reducing sugar assay or purified as describedbelow.

Variants above were produced by fermentation using standard protocols(TB-glycerol media containing a standard trace metal mix as described inF. William Studier, “Protein production by auto-induction inhigh-density shaking cultures”, Protein Expression and Purification, 41(2005) 207-234) and grown for 4 days at 30° C. before harvested).Supernatants of samples used for stress testing were inoculated from anovernight culture grown at 37° C. and subsequently fermented in 96-wellplate format (TB-glycerol media described above without calcium in thetrace metal mix for 4 days 30° C.).

Example 3: Purification of GH9 Endoglucanase Variants

The culture broth was centrifuged at 13′000 rpm (45 min, 18° C.,F12S-6×500 rotor) using a Sorval RC-6 plus centrifuge (ThermoFisherScientific). The supernatant was supplemented with (NH4)2SO4 to a finalconcentration of 0.8 M. The mixture was filtered using 0.2 μm bottle-toprapid flow filters (Nalgene). The mixture was loaded on a 50 mL PhenylSepharose High Performance (GE Healthcare, Uppsala, Sweden)pre-equilibrated with 20 mM Tris-HCl, pH 8.0 with 0.8 M (NH4)2SO4.Flowrate was set to 3 mL/min. After protein loading, the flow rate wasincreased to 5 mL/min and unbound or loosely bound protein was washedout by several column volumes of equilibration buffer. Elution wascarried out by step-wise increase of elution buffer (20 mM Tris-HCl, pH8.0). The target protein eluted during the (75-100%) elution step.Fractions of 8 mL were collected during the purification. The fractionswere evaluated using SDS-PAGE (NuPAGE, Invitrogen). Fractions elutingwith 20 mM Tris-HCl, pH 8.0 were pooled and desalted on a 350 mL G25desalting column pre-equilibrated with 20 mM Tris-HCl, pH 8.5. Thedesalted protein solution was applied on a 20 mL Source15Q columnpre-equilibrated with 20 mM Tris-HCl, pH 8.5 at 2 mL/min. Unbound orloosely bound proteins were washed using at least two column volumes ofequilibration buffer until a stable UV baseline was obtained. The flowrate was raised to 4 mL/min and elution was done by a linear NaClgradient using the elution buffer (20 mM Tris-HCl, pH 8.5+750 mM NaCl).3 mL fractions were collected during the purification. SDS-PAGE was usedto evaluate the fractions. Pure fractions were pooled and concentratedif necessary using Vivaspin 20 (10 kDa Cut-off, Sartorius). Proteinconcentration was determined using absorbance measurements at 280 nm.

Example 4: Detergent Stability Assay

GH9 endoglucanase (EG) activity (EC 3.2.1.4) was determined by reducingends on xanthan gum pre-treated with xanthan lyase using thecolorimetric assay developed by Lever (1972), Anal. Biochem. 47:273-279, 1972. Pre-treated xanthan gum is a modified form of the xanthansugar, where the terminal pyruvated mannose from side chains is removed(prepared according to Nankai et al. (1999) from the source Keltran).The GH9 mature parent endoglucanase and its variants cleave atbeta(1,4)-glucosyl bonds in the glucan backbone of pretreated xanthangum releasing glucans with a reducing end which can be determined byreaction with p-Hydroxybenzoic acid hydrazide (PAHBAH). The increase ofcolour is proportional to the enzyme activity under the conditions usedin the assay (e.g., Table 1) and used to estimate the residual activity(RA), half-life (T½) and the half-life improvement factor (HIF).

TABLE 1 Description of compositions and method steps of assays Stressassay: Detergent Persil Universal Gel Assay buffer (AB) 50 mM MOPS, 4 mMCaCl2, 0.01% Triton X-100, pH 7.0 Reference sample conditions 4° C. for17-20 hours Stress conditions 25° C. for 17-20 hours Activity assay:Substrate concentration 4 mg/mL modified xanthan gum Xanthan gumincubation 50° C. for 1 h PAHBAH solution 15 mg/mL 4-hydroxybenzoic acidhydrazide (PAHBAH), 50 g/L potassium sodium tartrate tetrahydrate, 20g/L NaOH PAHBAH development 95° C. for 10 min Method steps: 1. 30 μLenzyme sample (culture supernatant of variants or purified variants,10-150 ppm) are mixed with 270 μL detergent using magnetic stirringfor15 minutes in a micro titer plate (MTP). This plate is designated asthe ″stress MTP″. 2. 20 μL of the mixture is transferred to a new MTPand diluted 100-fold using a 2-step dilution (2 × 10-fold dilution). Thesample is diluted into assay buffer (AB): 50 mM MOPS, 4 mM CaCl₂, 0.01%Triton X-100, pH 7.0. This diluted MTP is the ″reference MTP″ and isstored at 4° C. over-night (at a time interval equal to that of thestress MTP below). 3. The stress MTP is incubated at 25° C. over-night(17-20 h). 4. After over-night incubation, the stress MTP is initiallymixed by magnetic stirring for 15 minutes 5. The stress MTP is thendiluted 100-fold as described for the reference MTP in step 2. 6. Toassess the enzymatic activity, 50 μL of diluted enzyme:detergent sample(from both reference and stress MTPs) is mixed with 50 μL 4 mg/mLmodified xanthan gum in PCR plates. The samples are then incubated at50° C. for 1 h. 7. Finally, the level of reducing ends is estimated byadding 75 μL PAHBAH solution (15 mg/mL PAHBAH, 50 g/L potassium sodiumtartrate tetrahydrate, 20 g/L NaOH) to all samples in the PCR plates.The samples are then incubated at 95° C. for 10 min. 8. After coolingdown to room temperature, the absorbance at 405 nm is measured. 9. Theresidual activity (RA) can calculated using the following formula:${{RA}\mspace{11mu}(\%)} = {\frac{{Abs}({Stress})}{{Abs}({Ref})} \times 100\%}$Abs(Stress): The absorbance at 405 nm of the sample in the stress MTP(incubated at 25° C. over-night) after subtracting relevant backgroundcontributions. Abs(Ref): The absorbance at 405 nm of the sample in thereference MTP (incubated at 5° C. over-night) after subtracting relevantbackground contributions. 10. Also, the half-lives for the degradationof each variant and parent endoglucanase are calculated using thefollowing formula (by applying 1st order kinetics for the degradation ofEG):${T\;{1/2}} = {- \frac{{\ln(2)} \times T}{\ln( \frac{{Abs}({Stress})}{{Abs}({Ref})} )}}$T: The incubation time. Abs(Stress) and Abs(Ref): See above in 9. 11.Half-life-improvement factors (HIFs) can then be calculated as:${HIF} = \frac{{{T1}/2},{variant}}{{{T1}/2},{wt}}$ T1/2, variant: Thehalf-life for a specific variant T1/2, wt (or wild-type): The half-lifefor EG wt (EG wild type), wherein said T1/2 wt is T1/2 of the matureparent endoglucanase with SEQ ID NO: 2.

The HIFs of the tested variants are shown in Tables 2-7 below: Allhalf-life values of the variants measured in culture supernatant werecalculated relative to the half-life of GH9 wild-type (mature parentendoclucanase with SEQ ID No: 2) measured as culture supernatant. Allhalf-life values of the variants measured as purified protein werecalculated relative to the half-life of GH9 wild-type (mature parentendoclucanase with SEQ ID No: 2) measured as purified protein. HIFs ofall variants were calculated based on the wild-type endoglucanase (SEQID NO: 2) incubated at the same detergent concentration and temperature(HIF of wild-type=1 in all tables, per definition).

TABLE 2 Variants of the mature parent GH9 endoglucanase (SEQ ID NO: 2)with corresponding half-life improvement factors (HIF) measured asculture supernatants Regions (as defined in example 1) Alteration HIF 4N285G 1.4 5 W333L 1.2 5 T353D 1.8 6 N558F 1.4 7 T633V 1.1 7 D635L 2.4 7D635M 1.2 7 D635T 1.1 7 F638Y 1.1 7 T639D 1.2 9 G994N 3.7

TABLE 3 Variants of the mature parent GH9 endoglucanase (SEQ ID NO: 2)with corresponding half-life improvement factors (HIF) measured asculture supernatants Regions (as defined in example 1) Alteration HIF 4K281T 1.3 6 N558NP >10.0 6 G563E 1.14 6 I575M >10.0 6 I575A 1.3 9 K921D1.4

TABLE 4 Variants of the mature parent GH9 endoglucanase (SEQ ID NO: 2)with corresponding half-life improvement factors (HIF) measured asculture supernatants Regions (as defined in example 1) Alteration HIF 2I125V 1.1 2 A126R 1.1 2 K130R 1.1 3 K213R 1.2 3 A221R 1.1 3 K228E 1.1 3K228I 1.2 3 G230F 1.2 3 G230L 1.2 3 G230A 1.2 3 G230H 1.2 3 G230N 1.1 3G230W 1.1 3 G230T 1.2 3 F231Y 1.2 3 F231N 1.1 3 V232R 1.1 3 V232G 1.1 3H235D 1.1 3 N240Q 1.2 3 G243K 1.1 3 G243R 1.2 3 A249N 1.1 4 A278S 1.1 4K281F 1.1 4 K281V 1.2 4 K281Y 1.1 4 K281H 1.1 4 K281Q 1.1 4 K281N 1.2 4K281W 1.1 4 N285L 1.9 4 N285M 1.8 4 N285S 1.2 4 N285P 1.6 4 N285T 1.1 4N285Y 1.8 4 N285H 2.4 4 N285K 1.4 4 N285D 1.6 4 N285W 1.3 4 N285R 1.5 4T292F 1.2 4 T292L 2 4 T292I 1.5 4 T292V 1.5 4 T292S 1.3 4 T292P 1.6 4T292Y 1.2 4 T292Q 1.4 4 T292N 1.1 4 T292K 1.2 4 T292D 1.3 4 T292G 1.4 4F297L 1.1 5 A346H 1.1 6 G556S 1.2 6 N558D 1.4 6 N558M 1.2 6 N558Q 1.3 6N558I 1.2 6 N558Y 1.1 6 N558H 1.1 6 A559N 1.7 6 A559F 1.2 6 A559M >10.06 A559P 1.5 6 A559Y 1.3 6 A559H 1.6 6 A559Q 1.4 6 A559D 1.5 6 A559R 1.16 A559G 1.1 6 A559I 1.1 6 A559S 1.2 6 S560P 1.3 6 S560K 1.2 6 S560G 1.46 S560D 1.3 6 T561P 1.6 6 T561E 1.1 6 T561Q 1.1 6 T561S 1.1 6 T561D 1.26 A564I 1.5 6 A564Y 1.2 6 A564H 1.1 6 A564Q 1.2 6 A564K 1.5 6 A564E 1.46 E565M 1.2 6 V567F 1.1 6 K568R 1.3 6 L569F 1.2 6 L569Y 1.2 6 L569D 1.26 L569E 1.3 6 P570F 1.3 6 P570L 1.6 6 P570I 1.8 6 P570M 2.1 6 P570V 2.66 P570S 4.9 6 P570T 5.4 6 P570A 4.4 6 P570Y 2.1 6 P570H 2.6 6 P570Q 5.16 P570N 3.3 6 P570K 4.2 6 P570E 1.6 6 P570W 1.7 6 P570R 3.6 6 P570G 1.46 I575D 1.5 6 I575E 3.7 6 I576F 1.8 6 I576M 1.2 6 I576P 3.3 6 D578R 1.16 Y579F 1.1 6 Y579W 2.1 6 V580L 1.2 6 D583M 1.1 6 Q589G 1.1 6 P590S 1.16 P590I 1.1 6 P590E 1.2 6 E591L 1.5 6 G592D 1.1 6 S593P 1.4 6 S593H 1.36 S593Q 1.3 6 S593N 1.5 6 S593K 1.2 6 S593D 1.3 6 S593E 1.3 6 S593R 1.37 S616D 1.2 7 K627L 1.8 7 K627M 2.2 7 K627V 2.4 7 K627S 1.1 7 K627T 1.77 K627Q 2.5 7 K627R 4.3 7 I630F 1.4 7 I630V 1.2 7 I630Y 1.2 7 D635A 1.37 D635P 1.1 7 D635N 1.3 7 D635K 1.3 7 D635E 1.4 7 D635G 1.1 7 D635W 1.17 S636L 1.6 7 S636M 1.9 7 S636A 1.4 7 S636H 1.4 7 S636Q 2.4 7 S636N 2 7S636K 1.8 7 S636R 1.3 7 F638I 1.7 7 F638V 1.4 7 F638T 1.2 7 F638L 1.1 7F638H 1.3 7 T639V 1.1 7 T639S 1.1 7 T639L 1.2 7 T639I 2.1 7 T639M 1.2 7T639A 2.2 7 T639E 1.8 7 T639W 1.3 7 T639G 2.1 7 Y641E 1.2 7 S642T 2.1 7S642N 1.3 7 N643D 2.0 7 N643H 2.1 7 N643T 1.2 7 T644F 1.1 7 A651P 1.5 8S810R 1.2 8 A811S 1.1 8 V812F 1.3 8 V812I 1.2 8 V812M 1.3 8 V812W 1.2 8V812R 1.1 8 N815V 1.1 8 N815Y 1.1 8 N815E 1.2 8 N815W 1.1 8 N815R 1.1 8S823Q 1.1 8 A824T 1.1 8 T825N 1.1 8 T825W 1.1 8 T825A 1.2 8 T825D 1.1 8V827I 1.2 8 V827M 1.5 8 V827S 1.2 9 T843V 1.1 9 D870F 1.2 9 D870L 1.3 9D870I 1.3 9 D870M 1.4 9 D870V 1.5 9 D870S 1.3 9 D870T 1.1 9 D870Y 1.1 9D870H 1.2 9 D870Q 1.3 9 D870N 1.5 9 D870K 1.4 9 D870E 1.4 9 D870W 1.1 9D870R 1.2 9 D870G 1.2 9 P871F 1.3 9 P871L 1.3 9 P871I 1.5 9 P871M 1.3 9P871V 1.3 9 P871S 1.3 9 P871T 1.4 9 P871A 1.3 9 P871Y 1.2 9 P871H 1.3 9P871Q 1.5 9 T872S 1.1 9 T872F 1.1 9 T872A 1.1 9 T872Y 1.2 9 T872H 1.6 9T872Q 1.5 9 T872N 1.5 9 T872K 1.4 9 T872D 1.4 9 T872E 1.5 9 T872W 1.5 9T872R 1.7 9 T872G 1.3 9 D873K 1.1 9 D873E 1.2 9 T874V 1.2 9 T874S 1.3 9T874P 1.1 9 T874A 1.2 9 T874H 1.1 9 T874Q 1.1 9 T874N 1.2 9 T874K 1.3 9V881Q 1.2 9 T883K 1.3 9 Y884H 1.8 9 A885F 5.4 9 A885Q 1.3 9 A885N 1.1 9T887L 1.1 9 T887I 1.1 9 T887S 1.6 9 T887H 1.1 9 T887R 1.1 9 K894E 1.1 9N920D 1.3 9 K921R 2.4 9 K921E 1.2 9 T932A 1.5 9 N933V 1.4 9 N933S 2.5 9Y934G 4.8 9 Y934M 1.2 9 Y934S 3.8 9 Y934A 3.7 9 Y934Q 4.2 9 Y934N 2.8 9Y934E 2.3 9 Y934W 1.5 9 Y934R 6 9 T935W 1.1 9 A937F 1.1 9 A937V 1.7 9A937S 1.1 9 A937T 1.4 9 A937Q 1.1 9 A937D 1.4 9 A937E 2.6 9 V938I 1.8 9K939I 1.8 9 K939V 3.4 9 D940E 1.1 9 N941S 2.4 9 N941H 1.6 9 N941D 1.3 9A942P 1.7 9 A942E 1.2 9 D943Y 1.1 9 D943H 1.2 9 R950V 1.2 9 R950H 1.2 9R950N 1.4 9 F952S 1.2 9 F952W 1.1 9 N953Y 1.7 9 G954L 1.4 9 Y960F 1.4 9A964N 1.2 9 A964C 1.2 9 N966P 1.1 9 N966C 1.5 9 G971A 1.1 9 Q974K 1.1 9Q974C 1.1 9 Q989I 1.1 9 Q991L 1.1 9 Q991I 1.2 9 Q991M 1.1 9 Q991V 1.2 9Q991T 1.3 9 Q991K 1.1 9 Q991C 1.1 9 S995I 1.1 9 S995V 1.1 9 S995Q 1.1 9S995R 1.1 9 S995C 1.3 9 G998V 1.1 9 G998A 1.1 9 S1006T 1.2 9 S1006A 1.19 S1006K 1.1 9 S1006R 1.2 9 Y1010W 1.1 9 L1011M 1.2 9 L1011S 1.3 9L1011A 1.4 9 L1011Q 1.1 9 L1011N 1.1 9 L1011D 1.2 9 L1011E 1.2 9 R1029N1.2 9 F1030M 1.1 9 K1031I 1.2 9 K1031S 1.4 9 K1031T 2.1 9 K1031H 1.2 9V1032G 1.3 9 K1035A 1.2 9 A1037E 1.4 9 A1037W 1.6 9 S1038L 1.1 9 S1038I1.2 9 L1040N 1.3 9 L1040E 1.1 9 G1041F 1.1 9 L1044F 1.1 9 L1044S 1.1 9L1044N 1.1 9 L1044W 1.1 9 P1045Q 1.2 9 P1045W 1.2

TABLE 5 Variants of the mature parent GH9 endoglucanase (SEQ ID NO: 2)with corresponding half-life improvement factors (HIF) measured asculture supernatants Alteration HIF A559N + Y579F 2.8 A564E + Y579F 1.1A559N + Y579W 3.2 G562P + Y579W 1.4 A564D + Y579W 2.1 A559N + Y579W +K99R 3.8 A559N + Y579W + K281R 3.4 K281R + A559N + Y579W 3.5 A559N +Y579W + 5616D 5.6 A559N + Y579W + 5636N 9 A559N + Y579W + A651P 4.2A559N + Y579W + K948E 2.6 A559N + Y579W + K1009E 2.4 A559N + Y579W +K627R 7.8 Y579W + K921R 3.2 A559N + Y579W + K921R 6.1 K99R + Y579W 1.6Y579W + A651P 2.7 Y579W + K948E 1.4 Y579W + K1009E 1.5 A559N + Y579W +Y934G 16.0 A559N + Y579W + K921R + Y934G 29.5 A559N + Y579W + K627M 6.4A559N + Y579W + K627R + S616D 18.1 A559N + Y579F + K627R 4.4 A559N +Y579W + K921R + A651P 9.9 A559N + Y579W + K921R + K627R 17.6 A559N +Y579W + K921R + S636K 5.9 A559N + Y579W + K921R + S616D 4.3 A559N +Y579W + K921R + S636N 8.9 A559N + Y579W + K921R + K627R + S636N 26.7A559N + Y579W + S636N + A651P 9.6 A559N + Y579W + S616D + A651P 9.4A559N + Y579W + S616D + S636K 6.5 A559N + Y579W + S616D + K921R + Y934G19.9 A559N + Y579W + A651P + K627M 10.6 A559N + Y579W + A651P + S636K8.2 A559N + Y579W + A651P + K627R + S636N 37.0 A559N + Y579W + A651P +S616D 9.0 A559N + Y579W + A651P + K921R + Y934G 21.1 S636N + Y934G 1.6S636N + K921R 1.4 S636N + K627R 2.4 S636N + Y579W 1.6 F638I + Y934G 3.0F638I + K921R 1.4 F638I + K627R 2.2 F638I + Y579W 1.3 K627R + K51Q 1.3K627R + K451S 1.4 K627R + A559N 1.5 K627R + Y579W 3.9 Y579W + Y934G 4.1A651P + F638I 1.2 P570Q + A651P 2.7 P570Q + K921R 1.6 P570Q + K627R 5.1P570Q + A559N 1.8 P570Q + Y579W 3.4 P570Q + F638I 1.2 P570K + Y579W 2.6P570K + F638I 1.5 P570T + A651P 2.6 P570T + S636N 3.4 P570T + Y934G 4.3P570T + F638I 1.4 P570T + K921R 1.5 P570T + K627R 3.9 P570T + A559N 1.8P570T + A885F 1.2 A885F + Y934G 1.5 A885F + K627R 1.2 A559N + Y579W +S636L 1.3 A559N + Y579W + F638I 2.1 A559N + Y579W + D870M 1.1 A559N +Y579W + S560P 4.1 A559N + Y579W + A564I 1.3 A559N + Y579W + P570N 2.8A559N + Y579W + P570K 5.2 A559N + Y579W + P570R 5.4 A559N + Y579W +P570A 4.1 A559N + Y579W + P570T 3.8 A559N + Y579W + P570S 3.9 A559N +Y579W + P570Q 3.3 A559N + Y579W + P570H 3.7

TABLE 6 Variants of the mature parent GH9 endoglucanase (SEQ ID NO: 2)with corresponding half-life improvement factors (HIF) measured aspurified samples Regions (as defined in example 1) Alteration HIF 6N558E 1.1 6 A559P 1.4 6 A559N 1.6 6 A559H 1.5 6 T561P 1.1 6 A564E 1.2 6P570A 7.0 6 P570Q 7.0 6 P570R 7.0 6 P570S 7.0 6 P570K 6.7 6 P570T 6.3 6P570N 2.9 6 Y579W 2.5 6 Y579F 1.3 6 T581M 1.2 7 S616D 1.4 7 K627R 2.1 7K627M 2.1 7 K627Q 1.2 7 S636N 1.8 7 S636Q 1.5 7 S636R 1.4 7 S636K 1.4 7S636M 1.3 7 S636H 1.1 7 F638I 1.6 7 F638L 1.2 7 N643D 1.5 7 A651P 1.3 7A651S 1.2 9 A885F 1.1 9 A885Q 1.1 9 K921R 4.2 9 Y934R 14 9 Y934G 6.2 9N966C 1.3 9 L1011A 1.1 9 K1031I 1.3

TABLE 7 Variants of the mature parent GH9 endoglucanase (SEQ ID NO: 2)with corresponding half-life improvement factors (HIF) measured aspurified samples Alteration HIF N558K + A559K + S560F + T561P + G562W3.8 A559N + P570A + Y579W 4.6 A559N + P570H + Y579W 3.5 A559N + P570K +Y579W 10.2 A559N + P570N + Y579W 3.4 A559N + P570Q + Y579W 4.5 A559N +P570R + Y579W 5.5 A559N + P570S + Y579W 5.0 A559N + P570T + Y579W 6.0A559N + S560P + Y579W 2.9 A559N + Y579W + A651P 1.6 A559N + Y579W +A651P + Y934G 6.5 A559N + Y579W + F638I 1.6 A559N + Y579W + K921R 1.4A559N + Y579W + S616D + K921R 7.4 A559N + Y579W + S636N 1.8 A559N +Y579F 1.7 A559N + Y579W 4.2 A559N + Y579W + K921R 5.8 A559N + Y579W +S616D 5.7 F638I + Y934G 1.8 K627R + S636N 1.5 K627R + Y934G 2.8 P570K +Y579W 3.1 Q416D + A559N + Y579W + S636N 3.9 S128X + A559N + Y579W +K627R 12.7 S128X + A559N + Y579W + S636N 7.6

Example 5: Half-Life Improvement Factors (HIF) of Endoglucanase Variantswith Mutations in Chelator-Induced Instability Regions and AdjacentRegions

Variants of the mature parent endoglucanase of SEQ ID NO: 2 wereprepared and purified as described above in Examples 2 and 3. For thepurposes of this example, variants were produced having mutations in atleast one chelator-induced instability region (regions 1, 2, 3, 4, 5, 6,7, 8, 9) and, optionally, in at least one adjacent region (regions 10,11, 12, 13, 14, 15, 16, 17, 18, 19). The in-detergent stability of thevariants was determined as described in Example 4 by measuring theenzymatic activity present in purified samples of the variants afterincubation with detergent and a protease. Incubation was performed usinga 90% or 95% concentration of Persil Universal Gel detergent (PUG), withincubation at a temperature of 25, 26, 28, or 30° C. and a variantincubation time ranging from 1½ hours to up to 720 hours.

Half-lives and HIFs were calculated as described above in Example 4. Incases where the difference in stability between wild-type and variantswas too big to accurately assess half-life for both wild-type andvariant using the same incubation time, the incubation time forwild-type and variant is different, e.g. 1 h for wild-type and up to 720h for the most stable variants.

Tables 8-12 below show the HIFs for the purified variants along withinformation on the test conditions (incubation temperature, detergentconcentration, incubation time) for each variant. HIFs of all variantswere calculated based on the wild-type endoglucanase (SEQ ID NO: 2)incubated at the same detergent concentration and temperature (HIF ofwild-type=1 in all tables, per definition).

TABLE 8 Variants of the mature parent GH9 endoglucanase (SEQ ID NO: 2)with corresponding half-life improvement factors (HIF) measured at atemperature of 25° C. Alterations compared to reference PUG (detergentIncubation (SEQ ID NO: 2) concentration) time (hrs) HIF A885F 90% PUG 161.1 S595L 90% PUG 16 1.1 T292A 90% PUG 18 1.1 F638I 90% PUG 16 1.5P570G + V837I 90% PUG 16 2.1

TABLE 9 Variants of the mature parent GH9 endoglucanase (SEQ ID NO: 2)with corresponding half-life improvement factors (HIF) measured at atemperature of 26° C. Alterations compared to reference PUG (detergentIncubation (SEQ ID NO: 2) concentration) time (hrs) HIF S593N 90% PUG 181.1 A957L 90% PUG 18 1.1 A942P 90% PUG 18 1.2 K281R 90% PUG 18.15 1.2T932A 90% PUG 18 1.4 V938I 90% PUG 18 1.4 N933S 90% PUG 18 1.5 A937E 90%PUG 18 1.6 Y884H 90% PUG 18 1.9 N941S 90% PUG 18 2.1 F638I 90% PUG 18.52.2 Y579W + K451Q 90% PUG 18 2.3 K939V 90% PUG 18 2.5 T711S + Y579W 90%PUG 19.5 2.6 Y579W + K451S + T972K 90% PUG 18 2.8 K713R + Y579W 90% PUG19.5 2.9 A564I 90% PUG 18 2.9

TABLE 10 Variants of the mature parent GH9 endoglucanase (SEQ ID NO: 2)with corresponding half-life improvement factors (HIF) measured at atemperature of 28° C. Alterations compared to reference PUG (detergentIncubation (SEQ ID NO: 2) concentration) time (hrs) HIF Y579W + Q834E90% PUG 5.5 3.5 Y579W + E408D 90% PUG 5.5 3.9

TABLE 11 Variants of the mature parent GH9 endoglucanase (SEQ ID NO: 2)with corresponding half-life improvement factors (HIF) measured at atemperature of 30° C. Alterations compared to reference PUG (detergentIncubation (SEQ ID NO: 2) concentration) time (hrs) HIF A559N + Y579W +T639G 90% PUG 16 3.5 A559N + Y579W + T639I + R898Q 90% PUG 16 3.5K451S + K627R 90% PUG 16 3.8 Q416S + F638I 90% PUG 16 3.9 V4T + A559N +Y579W 90% PUG 16 3.9 F638I + K921R 90% PUG 16 4.0 Q416S + S636N 90% PUG16 4.1 S636N + K921R 90% PUG 16 4.1 Y579W + S636N 90% PUG 16 4.1 Y55M +K627R + Y1042N 90% PUG 16 4.5 A559N + Y579W + S636L 90% PUG 16 4.7H149X + A559N + K627R 90% PUG 16 5.2 A448W + A559N + Y579W 90% PUG 165.7 K627R + K921R 90% PUG 16 5.9 E53Y + N216R + K627R 90% PUG 16 6.1E53Y + K627R 90% PUG 16 6.3 K627R + F638I 90% PUG 16 6.5 A448E + Y934G90% PUG 16 6.7 A559N + Y579W + A651P + S835A 90% PUG 168 6.7 P570K +F638I 90% PUG 16 7.0 E53Y + N216R + Y934G 90% PUG 16 7.4 F20P + F638I90% PUG 16 7.4 K451S + A559N + Y579W + S636N 90% PUG 168 8.5 F20P +A559N + Y579W 90% PUG 16 8.7 K451S + A559N + Y579W + S636N 90% PUG 1689.0 E408S + A559N + Y579W 90% PUG 16 9.6 Y579W + K627R 90% PUG 16 9.9P570T + Y934G 90% PUG 16 11.4 K512P + P570K 90% PUG 168 11.6 P570Q +K627R 90% PUG 16 11.6 A559N + Y579W + K627R 90% PUG 16 11.7 A559N +Y579W + I602T + V603P 90% PUG 168 13.2 P570T + E408D 90% PUG 16 14.3F20P + K627R 90% PUG 16 16.1 A559N + Y579W + I602T + V603P + S616D 90%PUG 168 17.3 A559N + Y579W + I602T + V603P + S616D + K921R 90% PUG 16818.0 E408D + A651P 90% PUG 16 18.4 P570Q + Y579W 90% PUG 16 19.0 Y55M +A559N + Y579W + A651P + Y934G 90% PUG 168 19.6 K51Q + K627R + Y934G 90%PUG 16 19.8 E408D + A559N + P570A + Y579W + K921R 90% PUG 16 20.7E408D + K512P + A559N + Y579W + F638I + A885F + 90% PUG 16 20.7 K921RE408D + Q416S + A559N + Y579W + S616D + K921R + 90% PUG 16 20.7 Y934GF20N + K627R + S636N + A651P + K921R + Y934G 90% PUG 16 20.7 F20N +S616D + K627R + A651P + K921R + Y934G 90% PUG 16 20.7 F20N + S616D +K627R + S636N + A651P + Y934G 90% PUG 16 20.7 K627R + S636N + A651P +K921R + Y934G 90% PUG 16 20.7 P570K + K627R + F638I 90% PUG 16 20.7P570Q + E408D 90% PUG 16 20.7 P570R + K627R + S636N 90% PUG 16 20.7P570S + K627R + S636N 90% PUG 16 20.7 S616D + K627R + S636N + A651P +K921R + Y934G 90% PUG 16 20.7 A559N + S560P + Y579W + K627R 90% PUG 1621.3 E408D + A559N + Y579W + S636N + K921R + Y934G 90% PUG 16 21.7E408D + A559N + Y579W + S616D + K921R 90% PUG 16 22.2 K627R + A651P +Y934G 90% PUG 138 24.4 F20P + F638I 90% PUG 138 25.7 S616D + K627R +K921R + Y934G 90% PUG 138 25.7 F20N + E408D + A559N + Y579W + K921R +Y934G 90% PUG 16 25.7 E408D + Y934G 90% PUG 16 25.9 E408D + Q416D +A559N + Y579W + I602T + V603P + 90% PUG 16 26.0 S616D + K921R + Y934GF20P + E53Y + Q416D + A448E + K627R + Y934G 90% PUG 138 26.1 E408D +A559N + Y579W + S636N + K921R 90% PUG 16 26.1 V56M + K627R + Y934G 90%PUG 138 26.9 E53Y + Y55M + E408D + A559N + Y579W + S636N + 90% PUG 1627.8 K921R + Y934G K627R + F638I + Y934G 90% PUG 138 27.8 K51Q + K627R +Y934G 90% PUG 138 28.0 K451S + A559N + Y579W + I602T + V603P + S636N 90%PUG 168 28.1 S616D + K627R + A651P + K921R + Y934G + G994D 90% PUG 13828.4 F20N + S616D + K627R + S636N + Y934G 90% PUG 138 28.5 A448E +K627R + Y934G 90% PUG 138 28.5 E408D + K451S + A651P 90% PUG 16 28.6K627R + S636N + Y934G 90% PUG 138 28.7 K627R + K921R + Y934G 90% PUG 13828.9 S607C + S1038G 90% PUG 138 29.0 K627R + S636K + Y934G 90% PUG 13830.0 F20P + Q416D + A559N + Y579W + K627R 90% PUG 138 30.7 A559N +Y579W + K627R + Y934G 90% PUG 138 30.9 F20N + S616D + K627R + Y934G 90%PUG 138 30.9 F20N + E408D + A559N + Y579W + S616D + K921R + 90% PUG 1631.3 Y934G P570Q + Y579W 90% PUG 138 31.5 K627R + Q834E + Y934G 90% PUG138 31.6 Q416S + A448W + A559N + K627R + Y934G 90% PUG 138 31.9 P570K +K627R + Y934G 90% PUG 138 32.3 A559N + Y579W + S616D + K627R 90% PUG 13832.9 F20N + S616D + K627R + A651P + K921R + Y934G 90% PUG 138 33.4F20N + E408D + A559N + Y579W + K627R + K921R + 90% PUG 16 33.8 Y934GK512P + K627R + Y934G 90% PUG 138 33.8 S616D + K627R + A651P + K921R +Y934G 90% PUG 138 34.6 S616D + K627R + S636N + A651P + K921R + Y934G 90%PUG 138 34.9 F20P + A448E + K627R + Y934G 90% PUG 138 35.5 K627R +S636N + A651P + K921R + Y934G 90% PUG 138 36.1 F20N + E408D + Q416S +A559N + Y579W + S616D + 90% PUG 16 36.1 K921R + Y934G P570K + K627R +F638I 90% PUG 138 36.1 S616D + K627R + S636N + K921R + Y934G 90% PUG 13836.5 F20N + K627R + Y934G 90% PUG 138 36.9 P570T + K627R + Y934G 90% PUG138 36.9 F20P + E53Y + Q416D + K627R + Y934G 90% PUG 138 37.1 F20P +E53Y + K627R + Y934G 90% PUG 138 37.3 P570Q + K627R + Y934G 90% PUG 13838.1 P570K + Y579W + Y934Q 90% PUG 138 38.1 F20N + S616D + K627R +S636N + A651P + Y934G 90% PUG 138 39.4 P570S + K627R + S636N 90% PUG 13839.6 A559N + Y579W + S616D + K627R + S636N 90% PUG 138 40.1 S616D +K627R + S636N + A651P + Y934G 90% PUG 138 41.5 Q416D + P570K + Y579W 90%PUG 138 41.9 A559N + Y579W + S616D + K627R + A651P 90% PUG 138 42.2A559N + Y579W + S616D + K627R + A651P 90% PUG 168 42.2 P570K + Y579W +A651P 90% PUG 138 43.0 P570Q + K627R + A651P 90% PUG 138 43.0 P570R +K627R + S636N 90% PUG 138 43.5 F20N + S616D + K627R + K921R + Y934G 90%PUG 138 43.7 E408N + Q416D + K627R + S636N 90% PUG 138 44.4 F20P +A559N + Y579W + K627R 90% PUG 138 44.4 A559N + Y579W + S616D + K627R +S636N + A651P 90% PUG 168 44.5 P570K + Y579W + Y934G 90% PUG 138 44.5P570Q + K627R + Y934R 90% PUG 138 44.5 Q416S + P570K + Y579W 90% PUG 16844.7 A559N + Y579W + S616D + K627R + S636N + A651P 90% PUG 138 44.9F20N + A559N + Y579W + S616D + K627R + A651P 90% PUG 138 44.9 F20P +E53Y + A559N + Y579W + K627R 90% PUG 138 46.2 K627R + S636N + Q416D +E408N + Q416S 90% PUG 138 46.2 P570K + Y579W + K927R 90% PUG 138 46.6E408D + S636N + A651P + N441G 90% PUG 168 46.7 F20P + Q416D + K627R +Y934G 90% PUG 138 47.9 P570K + Y579W + Q416D 90% PUG 138 48.8 A559N +S560P + Y579W + K627R 90% PUG 138 49.9 F20N + K627R + S636N + K921R +Y934G 90% PUG 138 50.1 E408D + A559N + Y579W + I602T + V603P + S616D +90% PUG 16 50.1 K921R F20N + A559N + Y579W + S616D + K627R + S636N + 90%PUG 138 52.1 A651P K512P + P570K + Y579W 90% PUG 138 52.3 F20N + K627R +S636N + A651P + K921R + Y934G 90% PUG 138 52.4 E408D + S636N + A651P +I575V 90% PUG 168 52.7 F20N + S616D + K627R + S636N + S642N + K921R +90% PUG 138 52.9 Y934G P570K + Y579W + S636N 90% PUG 138 53.6 E408D +A559N + Y579W + K921R 90% PUG 16 54.1 A559N + P570S + Y579W + K627R 90%PUG 138 55.3 A559N + S560P + Y579W + K627R 90% PUG 138 55.5 E408S +A559N + Y579W + K627R 90% PUG 138 55.7 P570K + Y579W + K627M 90% PUG 13855.9 E408D + S636N + A651P + A924D 90% PUG 168 56.5 E408D + Y579W 90%PUG 16 56.5 E408N + Q416D + Y579W + K627R 90% PUG 138 57.3 A559N +P570T + Y579W + K627R 90% PUG 138 58.0 E408D + S636N + A651P + A688G 90%PUG 168 58.1 P570Q + E408D 90% PUG 138 58.4 E408D + S636N + A651P +V756H 90% PUG 168 58.5 E408N + K627R + S636N 90% PUG 138 58.8 E408D +Y579W + A937E + F20P + T887S + S636K 90% PUG 168 59.9 E408D + S636N +A651P + T883R + Q956Y 90% PUG 168 60.4 E408D + S636N + A651P + N216D 90%PUG 168 60.5 E408D + A559N + Y579W + S616D + K921R + Y934R + 90% PUG 13860.7 A937E + K939V F20N + E408D + Q416S + A559N + Y579W + S616D + 90%PUG 138 60.9 K921R E408D + Q416S + A559N + Y579W + S616D + K921R + 90%PUG 138 61.2 Y934G A559N + P570R + Y579W + K627R 90% PUG 138 61.8E408D + Y934G 90% PUG 138 61.8 F20P + E408D + Y579W + S636K + A937E 90%PUG 168 62.1 E408D + S636N + A651P + K720H 90% PUG 168 62.1 F20N +E408D + A559N + Y579W + K921R + Y934G + 90% PUG 168 63.2 T883R E408D +A559N + Y579W + S616D + K921R + Q298E 90% PUG 138 63.5 E408D + A559N +Y579W + S616D + K921R + Y934R + 90% PUG 138 63.7 A937E + K939V E408D +Y579W + A651P + I602T + Q416S + S560P 90% PUG 168 63.8 E408D + Y579W +A937E + F20P + S636K 90% PUG 168 63.8 E408D + A559N + Y579W + I602T +V603P + S616D + 90% PUG 138 64.9 K921R + Y934G + A885Q E408D + A559N +Y579W + K921R 90% PUG 168 65.1 K627R + S636N + Q416D + E408N + Y579W 90%PUG 138 66.2 E408D + K512P + A559N + Y579W + F638I + A885F + 90% PUG 13866.3 K921R E408D + Y579W + A651P + I602T + Q416S + V926P 90% PUG 16866.5 E408D + A559N + Y579W + S616D + K921R + Y934G + 90% PUG 138 66.6A937E + K939V E408D + A559N + P570A + Y579W + K921R 90% PUG 138 67.0E408D + A559N + Y579W + S616D + K921R + D635A 90% PUG 138 67.6 E408D +A559N + Y579W + S616D + K921R 90% PUG 138 67.7 S17A + F20P + Q416D +P570K + Y579W 90% PUG 138 68.0 E408D + Y579W + A937E + F20P + S560P 90%PUG 138 69.1 E408N + Q416D + P570K + Y579W 90% PUG 138 69.5 F20N +E408D + Q416S + A559N + Y579W + S616D + 90% PUG 138 70.3 K921R + Y934GA559N + Y579W + P570K + Y934R + Q416S 90% PUG 168 70.3 P570K + Y579W +Q416D + E408N + S17A + F20P 90% PUG 138 70.5 E408D + A559N + Y579W +I602T + V603P + S616D + 90% PUG 168 71.6 K921R + Y934G + Y884H E408D +A559N + Y579W + S616D + K921R + Y934G 90% PUG 138 71.9 A559N + P570Q +Y579W + K627R 90% PUG 138 72.1 E408D + Q416D + A559N + Y579W + K921R +Y934G 90% PUG 138 72.1 F20N + P570K + Y579W 90% PUG 138 72.9 A559N +Y579W + P570K + Y934R + Q416D 90% PUG 168 72.9 A559N + Y579W + P570K +Y934R + F20P 90% PUG 168 74.9 E408D + Y579W + A651P + I602T + Q416S +V756Y 90% PUG 168 75.4 E408D + A559N + Y579W + I602T + V603P + S616D +90% PUG 138 75.9 K921R + Y934G + A885F E408D + Y579W + A651P + I602T +Q416S + I403Y 90% PUG 168 76.1 E408D + A559N + Y579W + I602T + V603P +S616D + 90% PUG 138 76.2 K921R + Y934G + Q834E E408D + A559N + Y579W +S616D + K921R + S313D 90% PUG 138 76.2 E408D + K451S + A651P 90% PUG 13876.2 E408D + S636N + A651P + H182Y 90% PUG 138 76.6 E408D + Y579W +A937E + F20P + V603P 90% PUG 138 77.8 E408D + A559N + Y579W + K921R 90%PUG 138 77.9 Y579W + A937E 90% PUG 138 77.9 E408D + Q416D + A559N +Y579W + K627R + K921R + 90% PUG 138 78.0 Y934G E408D + S636N + A651P +K631R 90% PUG 138 78.2 A559N + Y579W + P570K + Y934R + K627R 90% PUG 16878.7 P570K + Y579W + Q416D + S17A + F20P + E408N + 90% PUG 138 79.1Q416D E53Y + Y55M + E408D + A559N + Y579W + S636N + 90% PUG 138 80.1K921R + Y934G E408D + A559N + Y579W + K921R + Y934G + T697G + 90% PUG138 80.1 F20N E408D + Y579W + A651P + I602T + Q416S + V881Q 90% PUG 16880.1 E408D + S636N + A651P + T999R 90% PUG 138 81.0 E408D + A651P 90%PUG 138 81.1 E408D + A559N + Y579W + S636N + K921R 90% PUG 138 81.6E408D + Q416D + A559N + P570R + Y579W + K921R + 90% PUG 138 82.3 Y934GE408D + Y579W + Q834E 90% PUG 138 82.5 F20N + E408D + A559N + Y579W +K921R + Y934G 90% PUG 138 82.7 A559N + Y579W + P570K + Y934R + A651P 90%PUG 138 83.2 E408D + S636N + A651P + F1048W 90% PUG 138 83.3 E408D +Y579W + A651P + I602T + Q416S + Y579W 90% PUG 168 83.9 E408D + A559N +P570Q + Y579W + K921R + Y934G 90% PUG 138 84.1 E408D + S636N + A651P +R880K 90% PUG 138 84.1 F20P + E408D + Y579W + A937E 90% PUG 138 84.2E408D + A559N + Y579W + I602T + V603P + S616D + 90% PUG 138 84.5 K921R +A1037E E408D + A559N + Y579W + S616D + K921R + T697G + 90% PUG 138 84.5A885Q E408D + Y579W + A937E + F20P + T883R 90% PUG 138 84.7 E408D +Y579W + A651P + I602T + Q416S + K627R 90% PUG 168 85.1 F20N + E408D +A559N + Y579W + K921R + Y934G + 90% PUG 138 85.3 T999R E408D + Y579W +A937E + F20P + Q834E 90% PUG 138 85.5 P570K + Y579W + Q416D + S17A +F20P + Y934R 90% PUG 138 87.1 E408D + S636N + A651P 90% PUG 138 87.6P570K + Y579W + Q416D + S17A + F20P + I602T 90% PUG 138 87.6 E408D +S636N + A651P + Q298E 90% PUG 138 88.0 E408D + Y579W + A651P + I602T +Q416S + A559N 90% PUG 168 88.1 E408D + S636N + A651P + V926P 90% PUG 13888.9 E408D + A559N + Y579W + I602T + V603P + S616D + 90% PUG 138 88.9K921R + Y934G + S636K E408D + A559N + Y579W + S636N + A651P + K921R +90% PUG 138 89.0 Y934G E408D + A559N + Y579W + S616D + K921R + V756Y 90%PUG 138 89.5 P570K + Y579W + Q416D + S17A + F20P + I602T + V603P 90% PUG138 89.5 E408D + A559N + Y579W + S636N + K921R + Y934G 90% PUG 138 90.6E408D + Y579W + A937E + F20P + S636K 90% PUG 138 90.8 E408D + Q416D +A559N + Y579W + I602T + V603P + 90% PUG 138 90.9 S616D + K921R + Y934G +A651P + A885F E408D + A559N + Y579W + I602T + V603P + S616D + 90% PUG138 91.1 K921R + Y934G + S17A E408D + A559N + Y579W + I602T + V603P +S616D + 90% PUG 138 91.5 K921R + Y934G E408D + A559N + Y579W + S616D +T697G + V756Y + 90% PUG 138 91.5 K921R E408D + Y579W + A651P + I602T +Q416S + S17A + 90% PUG 168 93.5 F20P F20N + E408D + A559N + Y579W +K627R + K921R + 90% PUG 138 93.9 Y934G A559N + Y579W + P570K + Y934R +E408D 90% PUG 168 94.3 E408D + Q416D + A559N + Y579W + K921R 90% PUG 16894.7 E408D + A559N + Y579W + K921R + Y934G + A937E + 90% PUG 138 95.0K939V E408D + S636N + A651P + V756H + T883R 90% PUG 168 95.0 E408D +Y579W + A937E + F20P + S17A + F20P 90% PUG 138 95.0 E408D + A559N +Y579W + I602T + V603P + S616D + 90% PUG 138 95.5 K921R F20N + E408D +A559N + Y579W + K921R + Y934G + 90% PUG 138 95.9 I302D E408D + Q416S +Y579W + I602T + A651P + R880K 90% PUG 138 96.7 F20P + E408D + Q416S +A559N + S560P + P570R + 90% PUG 168 97.7 Y579W + K627R + K921R + Y934RF20P + E408D + A559N + Y579W + K921R + Y934G 90% PUG 138 98.0 E408D +A559N + Y579W + I602T + V603P + S616D + 90% PUG 168 98.1 K921R + Y934G +A937E + K939V E408D + S636N + A651P + T883K 90% PUG 138 98.5 N18G +A71E + L226K + E408D + Y579W + I602T + 90% PUG 168 98.5 A651P + A688G +V756Y + T887S E408D + A559N + Y579W + I602T + V603P + S616D + 90% PUG138 101.3 K921R + Y934R + A937E + K939V E408D + A559N + Y579W + I602T +V603P + S616D + 90% PUG 138 102.0 K921R E408D + A559N + Y579W + S616D +K921R + T697G + 90% PUG 138 102.2 A651P F20N + E408D + A559N + Y579W +K921R + Y934G + 90% PUG 138 102.2 R880K A559N + Y579W + P570K + Y934R +E408N 90% PUG 138 102.9 E408D + Y579W + A651P 90% PUG 138 103.7 P570K +Y579W + Q416D + S17A + F20P + A651P 90% PUG 138 103.7 F20P + E408D +Q416D + A559N + S560P + P570R + 90% PUG 168 103.8 Y579W + K627R +S636N + F638I + K921R + Y934R E408D + A559N + Y579W + S616D + T697G +K921R 90% PUG 138 104.1 F20P + E408D + Y579W + S636K + T697G + A937E 90%PUG 168 105.4 E408D + A559N + Y579W + I602T + V603P + S616D + 90% PUG138 106.4 K921R + N933S + Y934R + A937E + V938I + K939V + N941S + A942PF20P + E408D + Y579W + A651P 90% PUG 138 106.9 K51Q + E408D + Y579W +A651P 90% PUG 138 109.7 A559N + Y579W + P570K + Y934R + Q683E 90% PUG138 109.9 F20P + E408D + Q416D + A559N + S560P + P570Q + 90% PUG 168109.9 Y579W + K627R + K921R + Y934G E408D + Q416D + A559N + Y579W +I602T + V603P + 90% PUG 168 110.9 S616D + A651P + K921R + Y934G E408D +A559N + Y579W + S616D + K921R + T697G + 90% PUG 138 111.8 V881T E408D +A559N + Y579W + S636N + Q683E + K921R + 90% PUG 138 112.7 Y934G E408D +S636N + A651P + V756Y 90% PUG 138 113.4 F20P + E408D + Y579W + S636K +A937E 90% PUG 168 113.9 E408D + S636N + A651P + A885Q 90% PUG 138 114.4E408D + S636N + A651P + V756H + T883K 90% PUG 168 118.9 F20P + E408D +Q416S + A559N + S560P + P570R + 90% PUG 168 121.5 Y579W + K627R +S636N + F638I + K921R + Y934G E408D + Y579W + I602T + A651P 90% PUG 138122.1 E408D + S636N + A651P + T697G + T883R 90% PUG 168 123.5 E408D +S636N + A651P + V756H + R880K 90% PUG 168 124.3 E408D + Y579W + A651P +I602T + R880K 90% PUG 168 124.5 F20N + E408D + A559N + Y579W + K921R +Y934G + 90% PUG 138 125.1 T697G + Y55M E408D + Q416D + A559N + Y579W +I602T + V603P + 90% PUG 138 125.4 S616D + K921R + Y934G S17A + F20P +E408D + Y579W + I602T + A651P + 90% PUG 168 127.3 F906A + Y934G F20N +E408D + A559N + Y579W + K921R + Y934G + 90% PUG 138 127.7 T887S E408D +S636N + A651P + V756H + Y579W 90% PUG 168 129.7 E408D + S636N + A651P +S313D 90% PUG 138 132.5 E408D + Y579W + A651P + I602T + V881T 90% PUG138 135.9 E408D + Y579W + A651P + I602T + T887S 90% PUG 138 138.8E408D + Y579W + I602T + F638N + A651P + R880K + 90% PUG 168 139.5K921R + Y934G E408D + S636N + A651P + T697G 90% PUG 138 140.2 E408D +S636N + A651P + V756H + I602T + V603P 90% PUG 168 140.7 E408D + A559N +Y579W + S616D + K921R + T697G + 90% PUG 138 141.8 I602T + V603P E408D +Q416D + A559N + Y579W + I602T + V603P + 90% PUG 138 142.7 S616D +K921R + Y934G + A651P + Y884H F20N + E408D + A559N + Y579W + K921R +Y934G + 90% PUG 138 143.9 V756Y E408D + S636N + A651P + V756H + A885Q90% PUG 168 144.7 S17A + F20P + E408D + Y579W + I602T + A651P + 90% PUG168 150.2 F906A + Y934G + S636K F20N + E408D + A559N + Y579W + K921R +Y934G + 90% PUG 138 150.6 T697G + T887K F20N + E408D + A559N + Y579W +K921R + Y934G + 90% PUG 138 151.1 T697G + A885Q E408D + Y579W + A651P +I602T + R880K 90% PUG 138 151.2 S17A + F20P + E408D + Y579W + I602T +A651P + 90% PUG 168 151.5 F906A + Y934G + V881Q I302D + E408D + Q416S +Y579W + I602T + A651P + 90% PUG 168 152.0 R880K + Y934Q S17A + F20P +E408D + Y579W + I602T + A651P + 90% PUG 168 153.5 F906A + Y934G + L888MS17A + F20P + E408D + Y579W + I602T + A651P + 90% PUG 168 154.1 F906A +Y934G + S560P I302D + E408D + Q416S + Y579W + I602T + A651P + 90% PUG168 154.9 R880K + K744Q + N848D + A868E F20P + S313D + E408D + Y579W +S636K + T697G + 90% PUG 168 155.5 T887K + A937E F20P + S313D + E408D +Y579W + S636K + T697G + 90% PUG 168 156.2 A937E + A885Q E408D + A559N +Y579W + S616D + K921R + T697G + 90% PUG 138 156.9 Y934G S17A + F20P +E408D + Y579W + I602T + A651P + 90% PUG 168 157.2 F906A + Y934G + T887KE408D + Y579W + A651P + I602T + V881Q 90% PUG 138 157.6 E408D + S636N +A651P + A688G + A283D 90% PUG 168 162.3 S17A + F20P + E408D + Y579W +I602T + A651P + 90% PUG 168 163.7 N905D + T825G A559N + Y579W + A688G +V756Y + K921R + Y934G 90% PUG 168 164.8 E408D + S636N + A651P + V756H +Y934Q 90% PUG 168 165.3 S17A + F20P + E408D + Y579W + I602T + A651P +90% PUG 168 166.2 F906A E408D + Q416S + Y579W + I602T + A651P + N848D +90% PUG 168 168.9 A868E E408D + S636N + A651P + A688G + D476R 90% PUG168 169.4 1302D + E408D + Q416S + Y579W + I602T + A651P + 90% PUG 168171.5 R880K + V756H S17A + F20P + E408D + Y579W + I602T + A651P + 90%PUG 168 172.3 F906A + Y934G + S636N E408D + Q416D + A559N + Y579W +I602T + V603P + 90% PUG 138 176.9 S616D + K921R + Y934G + A651P + T887SS17A + F20P + E408D + Y579W + I602T + A651P + 90% PUG 168 178.3 N848DE408D + Q416S + Y579W + I602T + A651P + V756Y 90% PUG 168 181.5 E408D +Q416S + Y579W + I602T + A651P + T697G 90% PUG 168 204.0

TABLE 12 Variants of the mature parent GH9 endoglucanase (SEQ ID NO: 2)with corresponding half-life improvement factors (HIF) measured at atemperature of 30° C. Alterations compared to reference PUG (detergentIncubation (SEQ ID NO: 2) concentration) time (hrs) HIF E408D + A559N +Y579W + K921R + Y934R + A937E + 95% PUG 168 72.0 K939V E408D + A559N +Y579W + K921R + Y934R + A937E + 95% PUG 168 74.7 V938I + K939V + N941S +A942P E408D + Y579W + A651P + I602T + Q416S 95% PUG 168 80.0 E408D +Y579W + A937E + F20P + Q683E 95% PUG 168 82.7 E408D + S636N + A651P +A283D 95% PUG 336 91.3 E408D + S636N + A651P + N18G 95% PUG 336 91.3E408D + Y579W + A651P + I602T + S17A + F20P 95% PUG 168 91.3 E408D +S636N + A651P + A346D 95% PUG 336 96.7 E408D + S636N + A651P + A71E +A120P + A186P + 95% PUG 168 102.4 D247N F20N + E408D + A559N + Y579W +Q683E + K921R + 95% PUG 168 112.7 Y934G E408D + Y579W + A651P + I602T +Y934Q 95% PUG 168 113.3 E408D + S636N + A651P + T872G 95% PUG 336 116.7E408D + Y579W + A651P + I602T + T883R 95% PUG 168 120.0 F20N + E408D +A559N + Y579W + T697G + K921R + 95% PUG 168 122.0 Y934G E408D + Y579W +A651P + I602T + Q416S + R880K + 95% PUG 336 125.3 N216D E408D + A559N +Y579W + I602T + V603P + S616D + 95% PUG 168 129.5 K921R + Y934G + Q683EE408D + Y579W + A651P + I602T + Q416S + R880K + 95% PUG 336 139.2 F20PE408D + S636N + A651P + N848D 95% PUG 336 141.7 E408D + Y579W + A651P +I602T + Q416S + R880K + 95% PUG 336 148.8 I302D E408D + Y579W + I602T +F638N + A651P + R880K + 95% PUG 672 152.9 K921R + Y934G + K51Q F20N +E408D + A559N + Y579W + K921R + Y934G + 95% PUG 168 156.0 T697G + K512PE408D + Y579W + A651P + I602T + Q416S + R880K + 95% PUG 336 156.9 Y884HN18G + A120P + E408D + S636N + A651P + A688G + 95% PUG 672 161.5 V756YE408D + S636N + A651P + V756H + S313D 95% PUG 750 161.7 N18G + A71E +A186P + E408D + Y579W + I602T + 95% PUG 672 161.7 A651P + A688G + V756YE408D + S636N + A651P + V756H + T697G 95% PUG 672 164.9 E408D + Y579W +A651P + I602T + T883R + V756Y 95% PUG 750 167.6 F20P + E408D + Y579W +S636K + T697G + A937E + 95% PUG 336 168.4 V926F E408D + A559N + Y579W +S616D + K921R + T697G + 95% PUG 672 168.9 I602T N216D + I302D + E408D +Q416S + Y579W + I602T + 95% PUG 672 170.3 A651P + A688G + R880K +V890R + N905D E408D + Y579W + I602T + F638N + A651P + N848D + 95% PUG168 172.6 R880K + K921R + Y934G N18G + N189K + E408D + A559N + Y579W +A688G + 95% PUG 750 175.7 T697G + V756Y + K921R + Y934G E408D + Y579W +I602T + F638N + A651P + V756Y + 95% PUG 672 181.0 R880K + K921R +Y934G + P570R S313D + E408D + Y579W + I602T + F638N + A651P + 95% PUG750 182.5 R880K + K921R + Y934G E408D + Y579W + A651P + I602T + T883R +N848D 95% PUG 750 183.1 E408D + Y579W + I602T + F638N + A651P + R880K +95% PUG 672 183.3 N905D + K921R + Y934G + A283D E408D + Y579W + I602T +F638N + A651P + R880K + 95% PUG 672 183.3 N905D + K921R + Y934G + A885QE408D + Y579W + I602T + F638N + A651P + R880K + 95% PUG 750 183.4N905D + K921R + Y934G S17A + F20P + E408D + Y579W + I602T + A651P + 95%PUG 750 183.7 F906A + Y934G + S313D E408D + Y579W + I602T + F638N +A651P + R880K + 95% PUG 672 183.8 K921R + Y934G + Y55D E408D + Y579W +I602T + F638N + A651P + R880K + 95% PUG 672 184.0 K921R + Y934G + N848DS313D + I302D + E408D + Y579W + I602T + F638N + 95% PUG 672 186.3A651P + R880K + K921R + Y934G N18G + A186P + E408D + Q416S + Y579W +I602T + 95% PUG 672 186.8 A651P + A688G + V756Y E408D + Y579W + A651P +I602T + T883R + A688G 95% PUG 750 188.3 E408D + Y579W + I602T + A651P +T697G + T883R 95% PUG 672 189.9 N18G + E408D + S636N + A651P + A688G +T697G + 95% PUG 750 190.9 V756Y N18G + E408D + A559N + Y579W + I602T +V603P + 95% PUG 672 196.2 S616D + T697G + V756Y + K921R + T999R F20N +A346D + E408D + K512P + A559N + Y579W + 95% PUG 672 197.8 I602T +S636N + A651P + T697G + V756Y + A824D + A885N + K921R + Y934G F20N +E408D + Q489P + K512P + A559N + Y579W + 95% PUG 672 198.0 I602T +A651P + T697G + V756Y + A885N + K921R + Y934G + A937E E408D + Y579W +I602T + F638N + A651P + R880K + 95% PUG 672 200.7 K921R + Y934G + V756YE408D + Y579W + I602T + F638N + A651P + R880K + 95% PUG 672 200.8N905D + K921R + Y934G + N848D S17A + F20P + E408D + Y579W + I602T +A651P + 95% PUG 672 202.0 A688G E408D + S636N + A651P + A688G + N848D +V881Q + 95% PUG 168 202.3 N905D F20N + E408D + K512P + A559N + Y579W +T697G + 95% PUG 336 202.3 K921R + Y934G + A651P F20P + Y55M + S313D +E408D + Y579W + S636K + 95% PUG 672 202.8 T697G + A937E I302D + E408D +Q416S + Y579W + I602T + A651P + 95% PUG 672 204.5 R880K + T697G N18G +A186P + E408D + Y579W + I602T + A651P + 95% PUG 750 204.5 A688G +V756Y + V881T S17A + F20P + E408D + Y579W + I602T + A651P + 95% PUG 672205.5 F906A + Y934G + A688G I302D + S313D + E408D + Q416S + Y579W +I602T + 95% PUG 672 206.3 A651P + A688G + R880K + T892V + N905D + Q912VE408D + S636N + A651P + T697G + S313D 95% PUG 672 206.6 F20P + S313D +E408D + Y579W + S636K + T697G + 95% PUG 672 207.3 A937E + V603P F20P +S313D + E408D + Y579W + S636K + T697G + 95% PUG 672 207.5 A937E + T887SE408D + Y579W + I602T + F638N + A651P + R880K + 95% PUG 672 207.9N905D + K921R + Y934G + A885N I302D + E408D + Q416S + Y579W + I602T +A651P + 95% PUG 750 212.4 A688G + R880K E408D + Y579W + I602T + F638N +A651P + V756Y + 95% PUG 168 214.2 R880K + K921R + Y934G E408D + Y579W +I602T + F638N + A651P + A688G + 95% PUG 672 214.9 K720H + R880K +K921R + Y934G I302D + E408D + Q416S + Y579W + I602T + A651P + 95% PUG672 215.5 A688G + W719R + R880K + N905D + F1048W K51Q + E408D + Y579W +I602T + F638N + A651P + 95% PUG 672 216.0 R880K + K921R + Y934G I302D +E408D + Q416S + Y579W + I602T + A651P + 95% PUG 672 222.1 A688G +W719R + R880K + N905D + F1048W I302D + E408D + Q416S + Y579W + I602T +A651P + 95% PUG 672 224.2 A688G + V881Q + K921R Y55M + A283D + S313D +E408D + Y579W + I602T + 95% PUG 672 224.3 F638N + A651P + K720H +V756Y + N848D + A869V + R880K + Q912V + K921R + Y934R F20P + S313D +E408D + V567P + Y579W + S636K + 95% PUG 672 224.6 A651P + T697G +N848D + T883R + A937E + Q956Y + F1048W F20P + S313D + E408D + Y579W +S636K + T697G + 95% PUG 672 226.1 A937E + T883C S313D + A346D + E408D +Y579W + I602T + F638N + 95% PUG 672 230.5 A651P + T697G + N848D +R880K + A885N + K921R + Y934G E408D + Y579W + I602T + F638N + A651P +R880K + 95% PUG 672 231.7 N905D + K921R + Y934G + A688G E408D + Y579W +I602T + F638N + A651P + V756Y + 95% PUG 672 232.3 R880K + K921R +Y934G + N848D I302D + E408D + Q416S + Y579W + I602T + A651P + 95% PUG672 232.9 A688G + V881Q + K921R F20P + S313D + E408D + Y579W + S636K +T697G + 95% PUG 672 235.3 A937E + A885N F20P + S313D + E408D + Y579W +S636K + T697G + 95% PUG 750 236.9 A937E + E53Y F20P + S313D + E408D +Y579W + S636K + T697G + 95% PUG 750 238.2 A937E + R880K S17A + F20P +E408D + Y579W + I602T + A651P + 95% PUG 750 240.0 F906A + Y934G + T697GY55M + A283D + S313D + E408D + Y579W + I602T + 95% PUG 672 243.1 F638N +A651P + K720H + V756Y + N848D + A869V + R880K + N905D + K921R + Y934RS313D + E408D + S560P + Y579W + I602T + F638N + 95% PUG 672 244.9A651P + W719R + V756Y + T825G + N848D + A869V + D870V + R880K + N905D +K921R + Y934G F20P + S313D + E408D + Y579W + S636K + T697G + 95% PUG 750247.1 A937E + T883V S313D + E408D + S560P + Y579W + I602T + F638N + 95%PUG 672 250.9 A651P + W719R + V756Y + N848D + A869V + D870V + R880K +K921R + Y934G I302D + E408D + Q416S + Y579W + I602T + A651P + 95% PUG672 253.3 A688G + R880K + N905D F20P + S313D + E408D + Y579W + S636K +T697G + 95% PUG 336 253.9 A937E E408D + Y579W + I602T + F638N + A651P +V756Y + 95% PUG 672 254.7 R880K + K921R + Y934G + S313D E408D + Y579W +I602T + A651P + A688G + N848D + 95% PUG 672 255.1 T883R S17A + F20P +N216D + I302D + E408D + Q416S + 95% PUG 672 257.1 Y579W + I602T +A651P + W719R + T883R + F906A + Y934G + Q956Y I302D + E408D + Q416S +Y579W + I602T + A651P + 95% PUG 672 257.7 A688G + R880K + N905D + K921RS313D + E408D + Y579W + I602T + F638N + A651P + 95% PUG 672 258.5T697G + R880K + K921R + Y934G I302D + E408D + Q416S + Y579W + I602T +A651P + 95% PUG 672 259.4 A688G + V881Q + N905D + K921R I302D + E408D +Q416S + Y579W + I602T + A651P + 95% PUG 672 263.5 A688G + V881Q +N905D + K921R E408D + Y579W + I602T + A651P + A688G + N848D + 95% PUG672 268.3 T883R + M980I S313D + E408D + Y579W + I602T + F638N + A651P +95% PUG 672 272.3 T697G + R880K + K921R + Y934G S313D + E408D + S560P +Y579W + I602T + F638N + 95% PUG 672 272.7 A651P + W719R + V756Y +N848D + A869V + D870V + R880K + N905D + K921R + Y934G N216Q + I302D +H311N + E408D + Q416S + Y579W + 95% PUG 672 275.1 I602T + A651P +A688G + T697G + W719R + R880K + N905D + Q912V F20P + S313D + E408D +Y579W + S636K + T697G + 95% PUG 750 285.8 N848D + T887K + A937E S313D +E408D + Y579W + I602T + F638N + A651P + 95% PUG 672 288.9 T697G +R880K + K921R + Y934G Y55M + A283D + S313D + E408D + Y579W + I602T + 95%PUG 672 289.4 F638N + A651P + W719R + V756Y + N848D + A869V + R880K +K921R + Y934R F20N + E408D + K512P + A559N + Y579W + I602T + 95% PUG 672290.1 A651P + T697G + V756Y + K921R + Y934G Y55M + A283D + S313D +E408D + Y579W + I602T + 95% PUG 672 291.9 F638N + A651P + W719R +V756Y + N848D + A869V + R880K + K921R + Y934R A283D + S313D + E408D +A559P + Y579W + I602T + 95% PUG 672 292.7 F638N + A651P + W719R +V756Y + N848D + A869V + D870V + R880K + N905D + K921R + Y934G F20P +I302D + S313D + E408D + D476R + Y579W + 95% PUG 672 293.3 S636K +T697G + V756Y + V881Q + T887K + F906A + A937E S313D + E408D + S560P +Y579W + I602T + F638N + 95% PUG 672 295.1 A651P + W719R + V756Y +N848D + A869V + D870V + R880K + N905D + K921R + Y934G F20P + I302D +S313D + E408D + D476R + Y579W + 95% PUG 672 299.4 S636K + T697G +V756Y + T887K + F906A + A937E S17A + F20P + N216D + A283D + H311N +E408D + 95% PUG 672 300.1 Y579W + I602T + A651P + A688G + T883R +F906A + Y934G + Q956Y Y55M + A283D + S313D + E408D + A559P + Y579W + 95%PUG 672 303.3 I602T + F638N + A651P + T697G + W719R + N848D + R880K +K921R + Y934G + A937E F20P + S313D + E408D + Y579W + S636K + T697G + 95%PUG 750 304.5 N848D + T887K + A937E + G1041R F20P + S313D + E408D +V567P + Y579W + S636K + 95% PUG 672 307.5 T697G + N848D + A937E + Q956YF20P + I302D + S313D + E408D + D476R + Y579W + 95% PUG 672 309.7 S636K +T697G + V756Y + V881Q + T887K + F906A + A937E I302D + E408D + Q416S +Y579W + I602T + A651P + 95% PUG 672 317.0 A688G + W719R + R880K +N905D + F1048W S17A + F20P + N216D + A283D + H311N + E408D + 95% PUG 672319.1 S560P + Y579W + I602T + A651P + A688G + T883R + F906A + Q912V +Y934G + Q956Y Y55M + A283D + S313D + E408D + Y579W + I602T + 95% PUG 672321.6 F638N + A651P + W719R + V756Y + N848D + A869V + R880K + N905D +K921R + Y934R F20P + S313D + E408D + Y579W + S636K + T697G + 95% PUG 672325.9 N848D + A937E + Q956Y F20N + Y55M + H311N + E408D + K512P +A559N + 95% PUG 672 338.4 Y579W + A651P + T697G + N848D + K921R + Y934GY55M + A283D + S313D + E408D + Y579W + I602T + 95% PUG 672 338.5 F638N +A651P + W719R + V756Y + N848D + A869V + R880K + K921R + Y934R Y55M +A283D + I302D + S313D + E408D + Y579W + 95% PUG 672 342.9 I602T +F638N + A651P + W719R + V756Y + N848D + A869V + R880K + K921R + Y934RY55M + A283D + S313D + E408D + Y579W + I602T + 95% PUG 672 345.3 F638N +A651P + W719R + V756Y + N848D + A869V + R880K + A885N + K921R + Y934RY55M + A283D + S313D + E408D + Y579W + I602T + 95% PUG 672 345.9 F638N +A651P + W719R + V756Y + N848D + A869V + R880K + N905D + K921R + Y934RY55M + A283D + I302D + S313D + E408D + Y579W + 95% PUG 672 348.9 I602T +F638N + A651P + W719R + V756Y + N848D + A869V + R880K + K921R + Y934RS313D + E408D + Y579W + I602T + F638N + A651P + 95% PUG 672 349.6T697G + N848D + R880K + K921R + Y934G F20P + I302D + S313D + E408D +D476R + Y579W + 95% PUG 672 351.2 S636K + A651P + T697G + W719R +V756Y + N848D + V881T + T887K + F906A + A937E F20P + S313D + E408D +Y579W + S636K + T697G + 95% PUG 672 353.7 N848D + A937E F20P + S313D +E408D + Y579W + S616G + S636K + 95% PUG 672 356.3 T697G + W719R +T883V + N905D + A937E Y55M + A283D + S313D + E408D + Y579W + I602T + 95%PUG 672 364.7 F638N + A651P + W719R + V756Y + N848D + A869V + R880K +N905D + K921R + Y934R F20P + S313D + E408D + Y579W + S636K + A688G + 95%PUG 672 365.7 T697G + V881Q + N905D + K921R + A937E I302D + E408D +Q416S + Y579W + I602T + A651P + 95% PUG 672 367.3 A688G + W719R +K720H + R880K + N905D + F1048W Y55M + A283D + I302D + S313D + E408D +Y579W + 95% PUG 672 367.5 I602T + F638N + A651P + W719R + V756Y +N848D + A869V + R880K + K921R + Y934R F20P + S313D + E408D + Y579W +S636K + A688G + 95% PUG 672 375.4 T697G + N905D + A937E F20P + I302D +S313D + E408D + D476R + Y579W + 95% PUG 672 376.0 S636K + T697G +W719R + V756Y + V881Q + T887K + F906A + A937E Y55M + A283D + I302D +S313D + E408D + Y579W + 95% PUG 672 387.3 I602T + F638N + A651P +W719R + V756Y + N848D + A869V + R880K + N905D + K921R + Y934R A283D +S313D + E408D + Y579W + I602T + F638N + 95% PUG 672 396.0 A651P +T697G + K720H + V756Y + N848D + A869V + R880K + A885Q + K921R + Y934GF20P + I302D + S313D + E408D + D476R + Y579W + 95% PUG 672 406.2 S636K +T697G + W719R + V756Y + V881Q + T887K + F906A + A937E F20P + S313D +E408D + Y579W + S636K + A688G + 95% PUG 672 409.6 T697G + W719R +A885N + N905D + A937E Y55M + A283D + I302D + S313D + E408D + Y579W + 95%PUG 672 410.3 I602T + F638N + A651P + W719R + V756Y + N848D + A869V +R880K + K921R + Y934R Y55M + A283D + S313D + E408D + Y579W + I602T + 95%PUG 168 411.7 F638N + A651P + K720H + V756Y + N848D + A869V + R880K +K921R + Y934R F20P + I302D + S313D + E408D + Y579W + S636K + 95% PUG 672412.1 A688G + T697G + W719R + N905D + A937E N216D + I302D + H311N +E408D + Q416S + Y579W + 95% PUG 672 424.0 I602T + A651P + A688G +W719R + A869V + R880K + V890R + N905D Y55M + A283D + I302D + S313D +E408D + Y579W + 95% PUG 672 426.7 I602T + F638N + A651P + W719R +V756Y + N848D + A869V + R880K + A885N + N905D + K921R + Y934R F20P +N216Q + I302D + S313D + E408D + V567P + 95% PUG 672 434.7 Y579W +S636K + A651P + T697G + W719R + N848D + T883R + N905D + A937E + Q956Y +F1048W F20P + K51Q + I302D + S313D + E408D + D476R + 95% PUG 672 440.1Q489P + A559N + Y579W + I602T + S636K + A651P + T697G + W719R + V756Y +N848D + T883R + T887K + F906A + A937E F20P + I302D + S313D + E408D +D476R + Y579W + 95% PUG 672 442.1 S636K + T697G + W719R + V756Y +V881Q + T887K + F906A + A937E N216Q + I302D + H311N + E408D + Q416S +Y579W + 95% PUG 672 450.7 I602T + A651P + A688G + T697G + W719R +R880K + N905D + Q912V F20P + I302D + S313D + E408D + Y579W + S636K + 95%PUG 672 452.1 A688G + T697G + W719R + A885N + N905D + A937E F20P +I302D + S313D + E408D + D476R + Q489P + 95% PUG 720 452.5 A559N +Y579W + I602T + S636K + A651P + T697G + W719R + V756Y + N848D + T883R +T887K + F906A + A937E F20P + I302D + S313D + A346D + E408D + D476R + 95%PUG 672 471.0 Q489P + Y579W + S636N + T697G + W719R + V756Y + A824D +N848D + V881Q + T887K + F906A + S928D + A937E + F1048W Y55M + A283D +S313D + E408D + Y579W + I602T + 95% PUG 672 471.7 F638N + A651P +Y690F + T697G + W719R + V756Y + R880K + K921R + Y934G + A937E Y55M +A283D + I302D + S313D + A346D + E408D + 95% PUG 720 472.1 Q489P +A559P + Y579W + I602T + V603P + F638N + A651P + T697G + W719R + K744H +A824D + N848D + R880K + V881T + F906A + Q912V + K921R + Y934G + A937E +K948R + Q956Y + F1048W F20P + I302D + S313D + E408D + Y579W + S636K +95% PUG 672 472.2 A688G + T697G + W719R + N905D + A937E N216D + S313D +E408D + D476R + Y579W + I602T + 95% PUG 720 492.4 V603P + F638N +A651P + A688G + T697G + W719R + V756H + T825G + N833D + A869V + R880K +T887K + K921R + S928D + Y934G + N941S + K948R F20P + I302D + S313D +E408D + D476R + Y579W + 95% PUG 672 498.5 S636K + A651P + T697G +W719R + V756Y + N848D + V881T + T887K + F906A + A937E F20P + I302D +S313D + E408D + D476R + Q489P + 95% PUG 672 505.4 Y579W + S636N +T697G + W719R + V756Y + A824D + V881Q + T887K + F906A + A937E + T999R +A1037E F20P + I302D + S313D + A346D + E408D + D476R + 95% PUG 672 507.2Y579W + S636N + T697G + W719R + V756Y + A824D + N848D + V881Q + T887K +F906A + A937E + T999R + F1048W N216Q + S313D + E408D + D476R + Y579W +I602T + 95% PUG 672 521.0 F638N + A651P + T697G + W719R + R880K +T887K + K921R + Y934G S17A + F20P + S313D + E408D + Y579W + I602T + 95%PUG 672 524.6 A651P + T697G + W719R + N848D + T883R + F906A + Y934G +Q956Y + T999R F20P + I302D + S313D + E408D + D476R + Y579W + 95% PUG 672526.8 I602T + S636K + T697G + W719R + V756Y + V881Q + T887K + F906A +A937E + T999R + F1048W F20P + I302D + S313D + A346D + E408D + D476R +95% PUG 672 531.0 Y579W + S636N + T697G + W719R + V756Y + N848D +T883R + F906A + S928D + A937E N216D + S313D + E408D + D476R + A564E +Y579W + 95% PUG 720 552.6 I602T + F638N + A651P + Y690F + T697G +W719R + V756H + N833D + A869V + R880K + V881T + T887K + K921R + S928D +Y934G + T999R F20P + I302D + S313D + E408D + D476R + Y579W + 95% PUG 672557.2 S636K +T697G + W719R + V756Y + N848D + A869V + V881Q + T887K +N905D + F906A + Q912V + A937E + T999R + F1048W F20P + I302D + S313D +E408D + D476R + Y579W + 95% PUG 672 558.6 S636K + T697G + W719R +V756Y + N848D + A869V + V881Q + T887K +F906A + Q912V + A937E + T999R +F1048W F20P + I302D + S313D + E408D + D476R + Q489P + 95% PUG 672 565.9Y579W + S636N + T697G + W719R + V756Y + A824D + N848D + V881Q + T887K +F906A + S928D + A937E Y55M + A283D + I302D + S313D + A386P + E408D + 95%PUG 720 588.5 Q489P + A559P + Y579W + I602T + V603P + F638N + A651P +T697G + W719R + Q834E + N848D + R880K + V881T + T892P + F906A + Q912V +K921R + Y934G + A937E + F1048W F20P + I302D + S313D + E408D + Q489P +Y579W + 95% PUG 720 59E8 I602T + S636N + T697G + W719R + V756Y + A824D +N848D + V881Q + T887K + N905D + F906A + S928D + A937E + T999R + A1037E +F1048W F20P + I302D + S313D + E408D + D476R + Y579W + 95% PUG 672 592.0I602T + V603P + S636K + T697G + W719R + V756Y + N848D + A869V + V881Q +T887K + F906A + Q912V + A937E + F1048W F20P + I302D + S313D + E408D +Y579W + I602T + 95% PUG 672 592.7 S636K + A688G + T697G + W719R +V756Y + V881T + N905D + A937E F20P + I302D + S313D + E408D + Q416S +D476R + 95% PUG 672 597.1 Q489P + Y579W + I602T + S636K + A651P +T697G + W719R + V756Y + A824D + N833D + N848D + T883R + T887K + F906A +A937E Y55M + A283D + I302D + S313D + E408D + A559P + 95% PUG 720 604.2Y579W + I602T + V603P + F638N + A651P + T697G + W719R + N833D + N848D +R880K + V881T + F906A + Q912V + K921R + Y934G + A937E + N941S + T999RF20P + A186P + I302D + S313D + E408D + D476R + 95% PUG 672 616.5 Q489P +Y579W + A599S + I602T + S636K + A651P + T697G + W719R + V756Y + N848D +T883R + T887K + F906A + A937E Y55M + A283D + I302D + S313D + E408D +A559P + 95% PUG 720 622.2 Y579W + I602T + V603P + F638N + A651P +T697G + W719R + N848D + A869V + R880K + V881T + F906A + Q912V + K921R +Y934G + A937E N216D + S313D + E408D + D476R + Y579W + I602T + 95% PUG720 622.4 V603P + F638N + A651P + A688G + T697G + W719R + V756H +R880K + T887K + K921R + S928D + Y934G + K948R F20P + A283D + I302D +S313D + E408D + Y579W + 95% PUG 672 624.4 I602T + S636K + A651P +A688G + T697G + W719R + V756Y + T887K + N905D + A937E F20P + I302D +S313D + A346D + E408D + D476R + 95% PUG 672 629.5 Y579W + I602T +T697G + W719R + N848D + V881Q + T887K + F906A + S928D + A937E + T999RF20P + K51Q + I302D + S313D + E408D + D476R + 95% PUG 672 639.6 Q489P +Y579W + I602T + S636K + A651P + T697G + W719R + V756Y +N848D + T883R +T887K + F906A + A937E F20P + I302D + S313D + A346D + E408D + D476R + 95%PUG 672 644.4 Q489P + Y579W + S636N + T697G + W719R + V756Y + A824D +N848D + V881Q + T887K + F906A + A937E + T999R F20P + I302D + S313D +E408D + D476R + Q489P + 95% PUG 672 655.2 Y579W + I602T + S636N +T697G + W719R + V756Y + A824D + N848D + V881Q + T887K + N905D + F906A +A937E + T999R + A1037E + F1048W F20P + Q289E + I302D + S313D + E408D +D476R + 95% PUG 720 658.0 Q489P + A559P + Y579W + I602T + S636N +T697G + W719R + V756Y + A824D + N848D + R880K + V881Q + T887K + N905D +F906A + S928D + Y934G + T999R + A1037E + F1048W F20P + K51Q + I302D +S313D + E408D + D476R + 95% PUG 672 658.2 Q489P + Y579W + I602T +S636K + A651P + T697G + W719R + V756Y + A824D + N848D + T883R + T887K +F906A + S928D + A937E + A1037E N216D + S313D + A346D + E408D + D476R +Q489P + 95% PUG 720 662.8 A559P + Y579W + I602T + F638N + A651P +A688G + T697G + W719R + V756H + R880K + T887K + K921R + S928D + Y934GF20P + I302D + S313D + A346D + E408D + D476R + 95% PUG 672 663.4 Q489P +Y579W + I602T + T697G + W719R + V756Y + N848D + V881Q + T887K + F906A +A937E N216D + S313D + E408D + D476R + Q489P + A559P + 95% PUG 720 679.1Y579W + I602T + F638N + A651P + A688G + T697G + W719R + V756H + Q834E +R880K + T887K + T892P + K921R + S928D + Y934G F20P + I302D + S313D +E408D + D476R + Q489P + 95% PUG 672 693.2 A559N + Y579W + I602T +S636K + A651P + T697G + W719R + V756Y + N848D + T883R + T887K + F906A +S928D + A937E F20P + I302D + S313D + E408D + D476R + Q489P + 95% PUG 672701.7 Y579W + I602T + S636K + A651P + T697G + W719R + V756Y + N848D +T883R + T887K + F906A + A937E F20P + I302D + S313D + E408D + Q416S +D476R + 95% PUG 672 719.6 Q489P + A559N + Y579W + I602T + S636K +A651P + T697G + W719R + V756Y + N848D + T883R + T887K + F906A + A937EF20P + I302D + S313D + E408D + D476R + Y579W + 95% PUG 672 769.2 I602T +S636N + T697G + W719R + V756Y + A824D + N848D + V881Q + T887K + F906A +S928D + A937E + T999R + F1048W

While at least one exemplary embodiment has been presented in theforegoing detailed description, it should be appreciated that a vastnumber of variations exist. It should also be appreciated that theexemplary embodiment or exemplary embodiments are only examples, and arenot intended to limit the scope, applicability, or configuration of thevarious embodiments in any way. Rather, the foregoing detaileddescription will provide those skilled in the art with a convenient roadmap for implementing an exemplary embodiment as contemplated herein. Itbeing understood that various changes may be made in the function andarrangement of elements described in an exemplary embodiment withoutdeparting from the scope of the various embodiments as set forth in theappended claims.

The invention claimed is:
 1. A detergent composition comprising anendoglucanase variant, comprising an alteration at one or more positionsin one or more chelator-induced instability region selected from thegroup of: i) region 7 corresponding to amino acids 612 to 660 of SEQ IDNO: 2 ii) region 1 corresponding to amino acids 95 to 105 of SEQ ID NO:2, iii) region 2 corresponding to amino acids 115 to 138 of SEQ ID NO:2, iv) region 3 corresponding to amino acids 210 to 251 of SEQ ID NO: 2,v) region 4 corresponding to amino acids 267 to 301 of SEQ ID NO: 2, vi)region 5 corresponding to amino acids 339 to 361 of SEQ ID NO: 2, vii)region 6 corresponding to amino acids 547 to 595 of SEQ ID NO: 2, viii)region 8 corresponding to amino acids 806 to 828 of SEQ ID NO: 2, andix) region 9 corresponding to amino acids 839 to 1042 of SEQ ID NO: 2,wherein said variant has at least 86% sequence identity to SEQ ID NO: 2.2. The detergent composition comprising an endoglucanase variantaccording to claim 1, wherein said chelator-induced instability regionhas one or more of the following features: i) in the presence of achelator is less conformationally stable than one or more or all of itsadjacent regions; and/or ii) in the presence of a chelator is moreexposed to said chelator than one or more or all of its adjacentregions; and/or iii) in the presence of a chelator is more accessible tosaid chelator than one or more or all of its adjacent regions; and/oriv) in the presence of a chelator is more conformationally dynamic thanone or more or all of its adjacent regions; and/or v) in the presence ofa chelator is more receptive to deuterium incorporation than one or moreor all of its adjacent regions.
 3. The detergent composition comprisingan endoglucanase variant according to claim 1, wherein said variantfurther comprises one or more alterations in one or more adjacentregions, wherein said one or more adjacent regions is selected from thegroup of: i′) region 10 corresponding to amino acids 1 to 94 of SEQ IDNO: 2, ii′) region 11 corresponding to amino acids 106 to 114 of SEQ IDNO: 2, iii′) region 12 corresponding to amino acids 139 to 209 of SEQ IDNO: 2, iv′) region 13 corresponding to amino acids 252 to 266 of SEQ IDNO: 2, v′) region 14 corresponding to amino acids 302 to 338 of SEQ IDNO: 2, vi′) region 15 corresponding to amino acids 362 to 546 of SEQ IDNO: 2, vii′) region 16 corresponding to amino acids 596 to 611 of SEQ IDNO: 2, viii′) region 17 corresponding to amino acids 661 to 805 of SEQID NO: 2, ix′) region 18 corresponding to amino acids 829 to 838 of SEQID NO: 2, and x′) region 19 corresponding to amino acids 1043 to 1055 ofSEQ ID NO:
 2. 4. The detergent composition comprising an endoglucanasevariant according to claim 1, wherein said variant has at least 87%sequence identity to SEQ ID NO:
 2. 5. The detergent compositioncomprising an endoglucanase variant according to claim 1, wherein saidalteration at one or more positions is selected from the group ofalterations in positions: 4, 17, 18, 20, 51, 53, 55, 56, 60, 63, 71, 79,87, 92, 99, 120, 125, 126, 130, 137, 182, 186, 189, 192, 213, 216, 221,226, 228, 230, 231, 232, 233, 235, 240, 243, 247, 249, 278, 279, 281,283, 285, 289, 292, 294, 298, 302, 311, 313, 333, 346, 353, 358, 386,387, 388, 390, 403, 408, 410, 416, 441, 448, 451, 471, 472, 476, 489,507, 512, 515, 538, 555, 556, 557, 558, 559, 560, 561, 562, 563, 564,567, 568, 570, 575, 578, 579, 580, 581, 583, 589, 590, 591, 592, 593,595, 598, 599, 602, 603, 605, 607, 609, 616, 627, 630, 631, 635, 636,638, 639, 640, 641, 642, 643, 644, 651, 676, 683, 688, 690, 694, 698,699, 706, 711, 713, 1719, 720, 744, 749, 754, 756, 760, 781, 786, 797,810, 811, 812, 815, 823, 824, 825, 827, 828, 833, 834, 835, 837, 843,848, 868, 869, 870, 871, 872, 873, 874, 880, 881, 883, 884, 885, 887,888, 890, 892, 894, 898, 905, 906, 912, 920, 921, 924, 926, 927, 928,932, 933, 934, 935, 937, 938, 939, 940, 941, 942, 943, 946, 948, 950,952, 953, 954, 956, 957, 960, 966, 971, 972, 980, 989, 991, 994, 995,998, 999, 1006, 1009, 1010, 1011, 1029, 1030, 1031, 1032, 1035, 1037,1038, 1040, 1041, 1042, 1044, 1045, and 1048 of SEQ ID NO: 2, whereinnumbering is according to SEQ ID NO:
 2. 6. The detergent compositioncomprising an endoglucanase variant according to claim 1, wherein saidalteration at one or more positions is selected from the group ofalterations in positions: 17, 20, 216, 283, 302, 311, 313, 408, 476,579, 602, 627, 636, 638, 651, 688, 697, 719, 756, 880, 881, 883, 887,906, 921, 934, 937, and 956, wherein numbering is according to SEQ IDNO:
 2. 7. The detergent composition comprising an endoglucanase variantaccording to claim 1, wherein said alteration at one or more positionsis selected from the group of alterations in positions: S17A, F20P,N216D, N216Q, A283D, I302D, H311N, S313D, E408D, D476R, Y579W, I602T,K627R, S636N, S636K F638I, F638N, A651P, A688G, T697G, W719R, V756Y,R880K, V881Q, T883R, T887K, F906A, K921R, Y934G, A937E, and Q956Y,wherein numbering is according to SEQ ID NO:
 2. 8. The detergentcomposition comprising an endoglucanase variant according to claim 1,wherein said endoglucanase variant has degrading activity on xanthan gumpretreated with xanthan lyase.
 9. The detergent composition comprisingan endoglucanase variant according to claim 1, wherein said variant hasan improved stability in a detergent composition compared to a parentendoglucanase having SEQ ID NO:
 2. 10. The detergent compositioncomprising an endoglucanase variant according to claim 1, wherein saidvariant has an improved half-life as compared to a parent endoglucanasehaving SEQ ID NO: 2, or as compared to a wild-type endoglucanase havingSEQ ID NO:
 2. 11. The detergent composition comprising an endoglucanasevariant according to claim 10, wherein said variant has a half-life of aleast about 1.5 hours when measured at a temperature of about 25° C. anda detergent concentration of about 90%.
 12. The detergent compositioncomprising an endoglucanase variant according to claim 1, wherein saidvariant has a half-life improvement factor (HIF) of >about 1.0.
 13. Amethod of laundering or dishwashing comprising the step of applying thedetergent composition according to claim 1 to a textile or hard surface.14. A method of degrading xanthan gum comprising the step of applyingthe detergent composition of claim 1 to xanthan gum.
 15. The detergentcomposition comprising an endoglucanase variant according to claim 1,wherein said variant has at least 99% sequence identity to SEQ ID NO: 2.16. The detergent composition comprising an endoglucanase variantaccording to claim 1, wherein said chelator-induced instability regionhas one or more of the following features: i) in the presence of achelator is less conformationally stable than one or more or all of itsadjacent regions; and/or ii) in the presence of a chelator is moreexposed to said chelator than one or more or all of its adjacentregions; and/or iii) in the presence of a chelator is more accessible tosaid chelator than one or more or all of its adjacent regions; and/oriv) in the presence of a chelator is more conformationally dynamic thanone or more or all of its adjacent regions; and/or v) in the presence ofa chelator is more receptive to deuterium incorporation than one or moreor all of its adjacent regions, wherein said endoglucanase variant hasendoglucanase EC 3.2.1.4 activity on xanthan gum pretreated with xanthanlyase, wherein said variant has an improved stability in a detergentcomposition compared to a parent endoglucanase having SEQ ID NO: 2, andwherein said variant has an improved half-life as compared to the parentendoglucanase having SEQ ID NO: 2, or as compared to a wild-typeendoglucanase having SEQ ID NO:
 2. 17. The detergent compositioncomprising an endoglucanase variant according to claim 1 wherein saidendoglucanase variant has endoglucanase EC 3.2.1.4 activity on xanthangum pretreated with xanthan lyase.